_:b8124708 . _:b437201989 . _:b437201341 . _:b437200794 "3"^^ . . _:b437201346 . . _:b437200805 "3"^^ . _:b437200945 . _:b437200804 "3"^^ . . . _:b437200807 "3"^^ . _:b437202005 . . _:b437201357 . _:b437200806 "3"^^ . . _:b437200801 "3"^^ . _:b437200486 . _:b437200332 . _:b437200800 "3"^^ . _:b437200803 "3"^^ . _:b437200802 "3"^^ . _:b437200813 "3"^^ . _:b437201283 . _:b437200812 "3"^^ . _:b437201956 . . _:b437200815 "3"^^ . _:b437201009 . . _:b437200814 "3"^^ . _:b437201846 . _:b437200809 "3"^^ . _:b437201663 . _:b437200808 "3"^^ . _:b437200811 "3"^^ . . _:b437200810 "3"^^ . _:b437201776 . . _:b437200821 "3"^^ . . . _:b437200820 "3"^^ . . _:b437200823 "3"^^ . . _:b437201866 . _:b437200822 "3"^^ . . _:b437200817 "3"^^ . _:b437201098 . _:b437200659 . . _:b437200816 "3"^^ . . _:b437200819 "3"^^ . _:b437201214 . _:b437201101 . . _:b8124676 . _:b437201765 . _:b437200818 "3"^^ . _:b437200764 . _:b8124677 . . . _:b437201585 . _:b437200829 "3"^^ . _:b8124678 . . _:b437200828 "3"^^ . . _:b8124679 . _:b437200831 "3"^^ . . _:b8124672 . _:b437200830 "3"^^ . . _:b8124673 . . _:b437200825 "3"^^ . . _:b8124674 . _:b437200957 . _:b437200824 "3"^^ . _:b8124675 . _:b437200827 "3"^^ . _:b8124684 . . _:b437200826 "3"^^ . _:b437201093 . . _:b8124642 "The entry of HIV-1 into target cells involves the sequential binding of the viral gp120 Env protein to the CD4 receptor and a chemokine co-receptor (>>1<<). CCR5 is the major co-receptor used by HIV-1, and is expressed on key T cell subsets that are depleted during HIV-1 infection including memory T cells (2)." . _:b8124685 . . . _:b437200837 "3"^^ . _:b8124686 . . _:b437200836 "3"^^ . _:b8124687 . _:b437200222 . _:b437200839 "3"^^ . _:b8124680 . _:b437200838 "3"^^ . . _:b8124681 . _:b437200833 "3"^^ . _:b8124682 . _:b437200832 "3"^^ . _:b8124683 . _:b437201119 . _:b437200835 "3"^^ . _:b437200215 . _:b8124692 . _:b437201843 . _:b437200834 "3"^^ . . . _:b437201038 . _:b8124693 . . _:b8124694 "For analysis of lymphocyte populations in peripheral blood, a further lymphoid gate (low side scatter) was also applied to exclude cells of monocytic origin (>>74<<). All antibodies used were fluorochrome conjugated and human specific, and obtained from BD Biosciences: CD45 (clone 2D1), CD19 (clone HIB19), CD14 (clone M\u03D5P9), CD3 (clone SK7), CD4 (clone SK3), CD8 (clone HIT8a), CCR5 (2D7). Gates were" . . _:b437200845 "3"^^ . _:b8124694 . _:b437201666 . _:b437200844 "3"^^ . _:b437201507 . _:b8124695 . _:b437200847 "3"^^ . _:b8124688 . _:b437200846 "3"^^ . _:b8124689 . _:b437200841 "3"^^ . _:b437202003 . _:b437200995 . _:b8124690 . _:b437201458 . _:b437200840 "3"^^ . . . _:b8124691 . . _:b437201253 . _:b437200843 "3"^^ . _:b437201540 . . _:b8124700 . . _:b437201330 . _:b437200842 "3"^^ . _:b8124701 . _:b437200853 "3"^^ . _:b8124702 . _:b437200852 "3"^^ . _:b8124703 . _:b437200855 "3"^^ . _:b8124696 . _:b437200854 "3"^^ . _:b437201607 . _:b8124697 . _:b437200849 "3"^^ . _:b8124698 . _:b437200848 "3"^^ . _:b8124699 . _:b437200851 "3"^^ . _:b8124708 . . _:b437200850 "3"^^ . _:b8124709 . . _:b437200842 . _:b437200861 "3"^^ . _:b437200821 . _:b8124710 . _:b437201665 . _:b437200860 "3"^^ . _:b8124711 . _:b437200863 "3"^^ . _:b8124704 . _:b437200862 "3"^^ . _:b8124705 . _:b437200857 "3"^^ . . _:b437200971 . _:b8124706 . . _:b437200856 "3"^^ . _:b8124707 . _:b437200859 "3"^^ . _:b8124716 . _:b437200858 "3"^^ . . . _:b8124717 . . _:b437200869 "3"^^ . _:b8124718 . _:b437200868 "3"^^ . _:b8124719 . _:b437200871 "3"^^ . _:b8124712 . . _:b437200870 "3"^^ . _:b437201086 . _:b8124713 . _:b437200865 "3"^^ . _:b8124714 . _:b437200864 "3"^^ . _:b437201747 . _:b8124715 . _:b437200341 . _:b437200867 "3"^^ . _:b437201746 . _:b8124724 . _:b437200866 "3"^^ . _:b8124689 "The assay is sensitive enough to detect single-nucleotide changes and has a linear detection range between 0.69 and 44% (>>19<<)." . _:b8124725 . _:b437200770 . _:b437200373 . _:b437200877 "3"^^ . _:b8124726 . _:b8124647 . _:b437200876 "3"^^ . _:b437200288 . . . _:b8124727 . _:b437200522 . _:b437200879 "3"^^ . _:b437201508 . _:b8124720 . _:b437200878 "3"^^ . . _:b8124721 . _:b437200873 "3"^^ . . _:b8124722 . _:b437200872 "3"^^ . _:b8124723 . . _:b437200838 . _:b437200875 "3"^^ . . _:b437200874 "3"^^ . . _:b437200885 "3"^^ . _:b437200241 . . _:b437200884 "3"^^ . _:b437201440 . _:b437200788 . . _:b437200887 "3"^^ . _:b437200263 . . _:b8124728 . _:b437200585 . _:b437200886 "3"^^ . _:b8124729 . _:b437200881 "3"^^ . _:b437201137 . . _:b437200638 . . . _:b437200880 "3"^^ . _:b437200485 . _:b437200883 "3"^^ . . _:b437200882 "3"^^ . _:b437200893 "3"^^ . _:b437200196 . _:b437200892 "3"^^ . . . . _:b437200895 "3"^^ . _:b8124643 . _:b437200894 "3"^^ . _:b437200889 "3"^^ . _:b437201332 . _:b437200888 "3"^^ . _:b437201013 . . _:b437200891 "3"^^ . _:b437200890 "3"^^ . _:b437201717 . . _:b437201578 . . _:b437201371 . _:b437201453 . _:b437200487 . . _:b437200470 . . _:b437201785 . _:b437200356 . . . _:b437201296 . _:b437200418 . _:b437201979 . _:b437201711 . _:b8124712 . _:b437201429 . . _:b437201501 . _:b437201683 . _:b437200716 . _:b437201295 . _:b437201433 . . . . . . . . _:b437201576 . . _:b437201854 . _:b8124671 . _:b8124671 . . . . _:b8124674 "These observations mimic the human infection, where depletion of CD4+ CCR5+ lymphocytes results from a combination of direct infection, systemic immune activation and the upregulation of CCR5 on thymocytes (>>36<<,37,38). Following infection, blood samples were collected from the mice every two weeks and analyzed for HIV-1 RNA levels, T cell subsets and the extent of CCR5 disruption. At 8\u201312 weeks post-infection, animals were euthanized and" . _:b437200370 . . . _:b8124641 "introduction" . _:b437201689 . _:b437201660 . _:b437201206 . _:b437200475 . . . _:b437201012 . _:b437200725 . _:b437201745 . _:b437200393 . _:b437201861 . . . . . . _:b437200856 . . _:b437200363 . . _:b8124713 "the presence of HIV-resistant CCR5-negative cells in mucosal tissues would both protect individual cells from infection, and also help to break the cycle of immune hyperactivation that may underlie much of the pathology of AIDS (>>57<<)." . . . _:b437201089 . _:b437200891 . _:b437200404 . _:b437201582 . _:b437200587 . . _:b437201006 . _:b437201359 . . . . _:b437201963 . . _:b437200669 . . _:b437201032 . . . . . _:b437200571 . . . . _:b437200266 . _:b437200771 . _:b8124678 "Changes in the ratio of CD4+ to CD8+ T cells in the peripheral blood are characteristic of progressive infection in AIDS patients (39,>>40<<). We therefore examined the CD4:CD8 ratio in the peripheral blood samples from individual mice, both pre- and post-infection." . . _:b437201691 . _:b437200806 . _:b437201635 . . . . _:b437200874 . _:b437200264 . _:b437201966 . . _:b437201571 . _:b437201365 . _:b437201739 . _:b437200705 . _:b437201774 . _:b437201413 "2"^^ . . _:b437201412 "2"^^ . . _:b437200246 . . _:b437200608 . _:b437201415 "2"^^ . _:b8124729 "Our findings indicate that the modification of only a minority of human HSC was sufficient to provide the same strong anti-viral benefit as was conferred by a complete CCR5\u039432 stem cell transplantation in a patient (>>9<<). This suggests that a partially modified autologous transplant, administered under only mildly ablative transplantation regimens, may also be effective, opening up the treatment to many more HIV-infected individuals. Finally, the" . . . _:b437200958 . _:b437201414 "2"^^ . . _:b437201409 "2"^^ . . _:b437201408 "2"^^ . . _:b437201025 . _:b437201264 . . _:b437201411 "2"^^ . . _:b437200377 . _:b437201410 "2"^^ . . . . _:b437201421 "2"^^ . . . . _:b437201326 . _:b437201420 "2"^^ . _:b437200750 . _:b437201423 "2"^^ . _:b437201422 "2"^^ . _:b437201764 . . _:b437201417 "2"^^ . . _:b437201416 "2"^^ . _:b437201419 "2"^^ . . _:b437200632 . _:b437201847 . _:b437201418 "2"^^ . _:b437201429 "2"^^ . _:b437201489 . _:b437201428 "2"^^ . _:b8124704 "Moreover, unlike small molecule, antibody, or RNAi-based treatments (>>44<<), ZFN treatment results in disruption of the CCR5 open-reading frame and thus has the potential to completely eliminate this important HIV-1 co-receptor from the surface of cells that are bi-allelically modified." . _:b437201426 . . _:b437201431 "2"^^ . . _:b437201430 "2"^^ . . . . _:b437201425 "2"^^ . _:b437201424 "2"^^ . . _:b437201427 "2"^^ . _:b437201426 "2"^^ . _:b437200352 . _:b437201437 "2"^^ . _:b437201436 "2"^^ . _:b437200643 . . . _:b8124678 . _:b437201410 . _:b437201439 "2"^^ . _:b437200953 . _:b437201857 . _:b437201438 "2"^^ . . _:b437201433 "2"^^ . _:b437201710 . _:b437200590 . _:b437201432 "2"^^ . _:b437201852 . . _:b437201435 "2"^^ . . . _:b437201434 "2"^^ . . . _:b437201445 "2"^^ . _:b437201395 . _:b437201444 "2"^^ . _:b437200901 . _:b437201447 "2"^^ . _:b437201536 . . _:b437201446 "2"^^ . . _:b437201441 "2"^^ . . _:b437201440 "2"^^ . _:b437200626 . _:b437201443 "2"^^ . _:b437201442 "2"^^ . . _:b437201453 "2"^^ . _:b437200330 . . _:b437201553 . _:b437201452 "2"^^ . . _:b437201455 "2"^^ . . _:b437201245 . . . _:b437201454 "2"^^ . _:b437201449 "2"^^ . _:b437201381 . _:b8124701 "In this regard, the CCR5 co-receptor is a particularly attractive target because of the HIV-resistant phenotype of homozygous CCR5\u039432 individuals (3), who tolerate this genotype with minimal consequences (>>5<<\u20137). In the present study, we identified conditions that allowed the efficient disruption of the CCR5 gene in human CD34+ HSC, and further demonstrated that such modified HSC were able to generate CCR5-negative, HIV-resistant progeny in a" . . _:b437201448 "2"^^ . _:b8124652 "ZFNs comprise a series of linked zinc finger peptides that can be engineered to bind to DNA sequences in a highly specific manner, fused to an endonuclease domain (>>16<<). The concerted binding of two juxtaposed ZFNs on the DNA, followed by dimerization of the two endonuclease domains, results in a double stranded break (DSB) at the targeted DNA sequence. Such DSBs are rapidly repaired by cellular repair" . _:b437201451 "2"^^ . _:b437201053 . _:b437201450 "2"^^ . . _:b437201461 "2"^^ . . _:b437201460 "2"^^ . _:b437200645 . . _:b437201463 "2"^^ . _:b437200644 . _:b8124669 "NSG mice can be engrafted with human HSC (>>20<<) and thereby provide a rigorous readout of the hematopoietic potential of genetically modified HSC." . _:b437201462 "2"^^ . _:b437201825 . _:b437200647 . . _:b437201457 "2"^^ . _:b437200646 . _:b437201456 "2"^^ . _:b437200641 . _:b437201459 "2"^^ . _:b437200640 . _:b437200649 . _:b437201458 "2"^^ . _:b437201875 . _:b437200643 . _:b437201469 "2"^^ . _:b437200642 . _:b437200498 . _:b437201468 "2"^^ . _:b437200653 . . _:b437201471 "2"^^ . _:b437200652 . _:b437201470 "2"^^ . _:b437200655 . _:b437201412 . _:b437201465 "2"^^ . _:b437200654 . . _:b437201464 "2"^^ . _:b437200649 . _:b437201467 "2"^^ . _:b437200648 . _:b437201439 . . _:b437201466 "2"^^ . _:b437200651 . _:b437201719 . _:b437200932 . _:b437201477 "2"^^ . _:b437200973 . . _:b437200650 . _:b437200510 . _:b437201476 "2"^^ . _:b437200661 . . _:b437201479 "2"^^ . _:b437200660 . . _:b437201721 . . _:b437201478 "2"^^ . _:b437200663 . _:b437200252 . . _:b437201473 "2"^^ . _:b437200662 . . _:b437201472 "2"^^ . _:b437200657 . . _:b437201475 "2"^^ . _:b437200820 . _:b437201988 . _:b437200656 . _:b437201474 "2"^^ . _:b437200749 . _:b437200659 . _:b437200836 . _:b437201485 "2"^^ . _:b437200658 . _:b437201484 "2"^^ . _:b437200669 . _:b437201487 "2"^^ . _:b437201535 . _:b437200668 . . . _:b437201486 "2"^^ . _:b437200671 . . _:b437201481 "2"^^ . _:b437200670 . _:b437200326 . _:b437201480 "2"^^ . _:b437200665 . _:b437200333 . . _:b437201483 "2"^^ . _:b437200664 . _:b437200662 . . _:b437201329 . _:b437201482 "2"^^ . _:b437200667 . _:b437201493 "2"^^ . _:b437200666 . . . _:b437201210 . . _:b437201492 "2"^^ . _:b437200677 . _:b437201020 . _:b437201495 "2"^^ . _:b437200676 . _:b437200272 . . _:b437201494 "2"^^ . . _:b437200679 . _:b437200508 . _:b437200212 . _:b437201489 "2"^^ . _:b437200678 . . . _:b437201488 "2"^^ . . _:b437200673 . . _:b437201491 "2"^^ . _:b437200672 . _:b437200513 . _:b437201490 "2"^^ . _:b437200675 . _:b437201501 "2"^^ . _:b437200674 . . _:b437201500 "2"^^ . _:b437201383 . _:b437201081 . _:b437200949 . _:b437200685 . _:b437201149 . _:b437201503 "2"^^ . _:b437200684 . . _:b437201502 "2"^^ . _:b437200687 . . _:b437200805 . _:b437201497 "2"^^ . _:b437200903 . _:b437200686 . _:b437201496 "2"^^ . _:b437200681 . _:b437201674 . . . _:b437201499 "2"^^ . _:b437200680 . _:b437201498 "2"^^ . _:b437200683 . . _:b437201509 "2"^^ . _:b437200682 . _:b437201508 "2"^^ . _:b437200693 . . _:b437201511 "2"^^ . . _:b437200808 . _:b437200692 . _:b437201510 "2"^^ . _:b437200695 . _:b437201001 . . _:b437201505 "2"^^ . _:b437200694 . _:b437201302 . _:b437201504 "2"^^ . _:b437200689 . _:b437201291 . _:b437201507 "2"^^ . _:b437200688 . . _:b437200339 . _:b437201506 "2"^^ . _:b437200691 . _:b437200981 . _:b437201517 "2"^^ . _:b437200690 . _:b437201516 "2"^^ . _:b437200701 . _:b437200190 . . _:b437200978 . _:b437201519 "2"^^ . _:b437200700 . . _:b437201622 . _:b437201518 "2"^^ . _:b437200703 . _:b437200304 . _:b437201513 "2"^^ . _:b437200702 . _:b8124687 "Cg-Prkdc scid Il2rg tm1Wj/SzJ (NOD/SCID/IL2r\u03B3null, NSG) mice (>>71<<) were obtained from Jackson Laboratories (Bar Harbor, ME)." . _:b437200443 . _:b437201512 "2"^^ . _:b437200299 . _:b437201355 . . _:b437200697 . _:b437201515 "2"^^ . _:b437200696 . _:b437201267 . _:b437201514 "2"^^ . _:b437200699 . . _:b437201525 "2"^^ . _:b437200698 . _:b437200226 . _:b8124710 . . _:b437201524 "2"^^ . . _:b437200709 . . _:b437201298 . _:b437201806 . . _:b437201527 "2"^^ . _:b437200708 . . . . _:b437201526 "2"^^ . _:b437200233 . _:b437200232 . . _:b437200711 . _:b437200947 . _:b437201521 "2"^^ . _:b437200710 . _:b437201742 . _:b437201520 "2"^^ . _:b437200705 . . . . _:b437201523 "2"^^ . _:b437200704 . _:b437201268 . . _:b437201522 "2"^^ . _:b8124663 . _:b437200707 . . _:b437200837 . _:b437201533 "2"^^ . _:b437200706 . . _:b437201542 . _:b437201532 "2"^^ . _:b437200717 . _:b437201535 "2"^^ . _:b437200716 . . _:b437201534 "2"^^ . _:b437200719 . _:b437201529 "2"^^ . _:b437200718 . _:b437201921 . _:b437201528 "2"^^ . _:b437200713 . . _:b437201531 "2"^^ . _:b437200712 . . _:b437201530 "2"^^ . _:b8124654 . _:b437200715 . _:b437201215 . _:b437201541 "2"^^ . _:b437200714 . _:b437200389 . _:b437200223 . _:b437201540 "2"^^ . _:b437200725 . . _:b437201543 "2"^^ . _:b437200724 . _:b437201484 . _:b437201199 . _:b437201542 "2"^^ . _:b437200727 . . _:b437200839 . _:b437201537 "2"^^ . _:b437200677 . _:b437201392 . _:b437200726 . _:b437201536 "2"^^ . _:b437200721 . _:b437201539 "2"^^ . _:b437200720 . _:b437201538 "2"^^ . _:b437200723 . _:b437200740 . _:b437201549 "2"^^ . _:b437200722 . _:b437201207 . . _:b437200240 . _:b437201548 "2"^^ . . _:b437200733 . _:b437201551 "2"^^ . _:b437200732 . _:b437201938 . _:b437201550 "2"^^ . _:b437200735 . . _:b437201545 "2"^^ . _:b437200734 . _:b437200441 . _:b437200786 . . _:b437201544 "2"^^ . _:b437200729 . _:b437200401 . _:b437201547 "2"^^ . _:b437200728 . . _:b437201546 "2"^^ . _:b437201024 . _:b437200303 . _:b437200731 . . . _:b437201557 "2"^^ . _:b437200730 . _:b437200547 . _:b437201496 . _:b437201870 . _:b437201556 "2"^^ . _:b437200741 . _:b437200968 . _:b437201559 "2"^^ . _:b437200740 . _:b437201558 "2"^^ . _:b437200743 . . . _:b437201553 "2"^^ . . _:b437200742 . _:b437201552 "2"^^ . _:b437200737 . . _:b437201411 . . _:b437201555 "2"^^ . _:b8124701 . _:b437200736 . _:b437201628 . _:b437201554 "2"^^ . _:b437200739 . _:b437201519 . . _:b437201209 . _:b437201565 "2"^^ . _:b437200738 . _:b437200274 . _:b437201564 "2"^^ . _:b437200749 . _:b437201567 "2"^^ . _:b437200748 . . _:b437201566 "2"^^ . _:b437200751 . _:b437200835 . _:b437201561 "2"^^ . _:b437200750 . . _:b437200194 . . _:b437200306 . _:b437201560 "2"^^ . _:b437201157 . _:b437200745 . . _:b437200812 . _:b437201563 "2"^^ . _:b437200744 . _:b437201867 . . _:b437201562 "2"^^ . _:b437200747 . . . _:b437201573 "2"^^ . . _:b437200746 . . _:b437201572 "2"^^ . _:b437200757 . _:b437201050 . _:b437200584 . _:b437201575 "2"^^ . _:b437200756 . _:b437201690 . _:b437201574 "2"^^ . _:b437200759 . _:b437200219 . . _:b437201569 "2"^^ . _:b437200758 . . _:b437200653 . _:b437201568 "2"^^ . . _:b437200753 . _:b437201571 "2"^^ . _:b437200752 . . . _:b437201570 "2"^^ . _:b437200755 . . . _:b437201753 . _:b437201581 "2"^^ . _:b437200754 . _:b437201287 . _:b8124725 "It has been suggested that low level virus replication continues in certain sanctuary sites, even in well-controlled patients on ART (>>43<<,68), and this could provide a low level of selection, although drug intensification trials have not provided evidence of any such ongoing HIV-1 replication (69)." . _:b437201995 . _:b437201580 "2"^^ . _:b437200765 . . _:b437201583 "2"^^ . _:b437200764 . . _:b437201582 "2"^^ . _:b437200767 . _:b437201373 . _:b437201577 "2"^^ . _:b437200766 . . _:b437201143 . _:b437201576 "2"^^ . _:b437200761 . . _:b437201579 "2"^^ . _:b437200760 . _:b437201487 . _:b437201578 "2"^^ . _:b437200763 . . _:b437201589 "2"^^ . _:b437200762 . . . _:b437201588 "2"^^ . _:b437200773 . _:b437200360 . _:b437201591 "2"^^ . _:b437200772 . . _:b437201590 "2"^^ . . _:b437200775 . . . _:b437201398 . . _:b437201585 "2"^^ . _:b437200774 . _:b437201018 . _:b437200313 . _:b437201584 "2"^^ . _:b437200769 . _:b437201384 . _:b437201587 "2"^^ . _:b437200768 . _:b437200843 . . _:b437201586 "2"^^ . _:b437200771 . . _:b437201597 "2"^^ . _:b437200770 . _:b437200980 . _:b437201596 "2"^^ . _:b437200781 . _:b437201563 . _:b437201599 "2"^^ . _:b437200780 . _:b437200488 . _:b437201598 "2"^^ . . _:b437200783 . . _:b8124664 . _:b437201593 "2"^^ . _:b437200782 . _:b437200758 . . _:b437201592 "2"^^ . _:b437200777 . . _:b437201595 "2"^^ . _:b437200776 . _:b437201594 "2"^^ . _:b437200779 . _:b437201084 . . _:b437201605 "2"^^ . _:b437200609 . _:b437200778 . . _:b437200533 . _:b437201604 "2"^^ . . . _:b437200789 . _:b437200938 . _:b437201607 "2"^^ . _:b437201408 . _:b437200788 . _:b437201606 "2"^^ . _:b437201936 . _:b437200791 . . _:b437200268 . _:b437200396 . _:b437201601 "2"^^ . _:b437200790 . _:b437201297 . _:b437201600 "2"^^ . _:b437200785 . _:b437201603 "2"^^ . _:b437200784 . _:b437201495 . _:b437201602 "2"^^ . _:b437200787 . _:b8124706 . _:b437201613 "2"^^ . . _:b437200786 . . _:b437201612 "2"^^ . _:b437200797 . . _:b437201615 "2"^^ . _:b437200796 . _:b437200204 . _:b437201614 "2"^^ . _:b437200799 . . _:b437201609 "2"^^ . _:b437200798 . _:b437201788 . _:b437200974 . _:b437201608 "2"^^ . _:b437200793 . _:b437200192 . _:b437201611 "2"^^ . _:b437200792 . _:b437201589 . . _:b437201610 "2"^^ . _:b437200795 . _:b8124718 "In addition, the experience of autologous HSC transplantation in gene therapy for patients with genetic diseases such as ADA-SCID (59\u201360), chronic granulomatous disease (61) and X-linked adrenoleukodystrophy (>>62<<), is that non-myeloablative conditioning can be used to facilitate engraftment of gene-modified autologous HSC with minimal associated toxicity." . _:b437200235 . _:b437201858 . . _:b437201621 "2"^^ . _:b437200794 . _:b437201620 "2"^^ . _:b437200805 . _:b437201165 . . . _:b437201623 "2"^^ . _:b437200804 . . . _:b437201622 "2"^^ . . _:b437200684 . _:b437200807 . . _:b437201617 "2"^^ . . _:b437200806 . _:b8124689 . _:b437201616 "2"^^ . _:b437200801 . _:b437201949 . _:b8124688 . _:b437201619 "2"^^ . _:b437200800 . . _:b437201618 "2"^^ . _:b437200803 . _:b8124690 . . . . _:b437201629 "2"^^ . _:b437200802 . _:b437201628 "2"^^ . _:b437200813 . . _:b8124692 . . _:b437201417 . _:b437201631 "2"^^ . _:b437200812 . . _:b437201630 "2"^^ . _:b437200815 . _:b437201692 . . _:b8124677 . _:b437201625 "2"^^ . _:b437201282 . _:b437200814 . _:b437200187 "61"^^ . _:b437201624 "2"^^ . _:b437200809 . . _:b437200190 "30"^^ . _:b437201627 "2"^^ . _:b437200808 . . _:b437201626 "2"^^ . _:b437200189 "40"^^ . _:b437200811 . _:b437201637 "2"^^ . _:b437200810 . _:b437200188 "50"^^ . _:b437201961 . _:b437201636 "2"^^ . _:b437200821 . . . _:b437201639 "2"^^ . _:b437200820 . _:b437200999 . . _:b437201638 "2"^^ . _:b437200823 . . _:b8124702 . . _:b437201633 "2"^^ . _:b437200822 . _:b437201632 "2"^^ . _:b437200817 . . . _:b437201635 "2"^^ . . _:b437200816 . _:b437201634 "2"^^ . _:b437200819 . _:b437200624 . . _:b437201673 . _:b437201645 "2"^^ . . _:b437200186 "71"^^ . _:b437200818 . _:b437201069 . _:b437200956 . _:b437201644 "2"^^ . _:b437200829 . _:b437201647 "2"^^ . _:b437200828 . _:b437201646 "2"^^ . _:b437200831 . _:b437201641 "2"^^ . _:b437200830 . _:b8124681 . _:b437201640 "2"^^ . _:b437200825 . _:b437200406 . . _:b8124728 . _:b437201643 "2"^^ . _:b437200824 . _:b437201679 . _:b8124683 . . _:b437201642 "2"^^ . _:b437200827 . _:b8124682 . _:b437201653 "2"^^ . _:b437200826 . _:b437201780 . _:b437201652 "2"^^ . . _:b437200837 . . _:b437201655 "2"^^ . _:b437200836 . _:b437201624 . _:b437200458 . _:b437201654 "2"^^ . _:b437200839 . . _:b8124686 . . _:b437201649 "2"^^ . . _:b437200838 . _:b437200862 . _:b437200411 . _:b437201514 . _:b437201648 "2"^^ . _:b437200833 . _:b437201779 . _:b8124671 . . _:b437201651 "2"^^ . _:b437200832 . _:b437200875 . _:b437201621 . _:b437201650 "2"^^ . . _:b437200835 . . _:b437200191 "27"^^ . _:b437201868 . _:b437201661 "2"^^ . _:b437201597 . _:b437200834 . _:b437201660 "2"^^ . _:b437200845 . . _:b437201644 . _:b437200189 . _:b437201663 "2"^^ . _:b437200844 . _:b437201218 . . . _:b437201662 "2"^^ . _:b437200847 . _:b437200679 . _:b437201657 "2"^^ . . . _:b437200846 . _:b437200194 "25"^^ . _:b437200195 "23"^^ . _:b437201656 "2"^^ . . _:b437200841 . _:b437201074 . _:b437200193 "26"^^ . . _:b437201659 "2"^^ . _:b437200840 . _:b437201372 . _:b437200192 "26"^^ . _:b437201320 . _:b437201658 "2"^^ . _:b437200843 . . _:b437201669 "2"^^ . _:b437200842 . . _:b437200197 "21"^^ . _:b437201668 "2"^^ . _:b437200853 . _:b437201730 . _:b437200196 "23"^^ . _:b437201671 "2"^^ . _:b437200199 "20"^^ . _:b437201887 . _:b437200852 . _:b437200703 . . _:b437201670 "2"^^ . _:b437200198 "20"^^ . _:b437200855 . _:b437201665 "2"^^ . _:b437200201 "20"^^ . _:b437200854 . . _:b437201664 "2"^^ . _:b437200200 "20"^^ . _:b437200849 . _:b437201085 . _:b437200203 "19"^^ . _:b437201667 "2"^^ . . _:b437200848 . _:b437200507 . _:b437200202 "19"^^ . _:b437201666 "2"^^ . _:b437200851 . . _:b437200205 "18"^^ . _:b437201677 "2"^^ . _:b437200850 . . _:b437200204 "18"^^ . . _:b437200941 . _:b437201676 "2"^^ . _:b437200861 . . _:b437201430 . _:b437201679 "2"^^ . _:b437201929 . _:b437200860 . . . _:b437200206 "18"^^ . . _:b437201678 "2"^^ . _:b437200581 . . _:b437200863 . _:b437200207 "17"^^ . _:b437200870 . _:b437201673 "2"^^ . _:b437200302 . _:b437200862 . _:b437200208 "17"^^ . _:b437200734 . _:b437201672 "2"^^ . _:b437200209 "16"^^ . . _:b437200857 . _:b437200335 . _:b437201675 "2"^^ . _:b437200210 "16"^^ . _:b437200365 . _:b437200856 . _:b437201251 . _:b437201674 "2"^^ . _:b437200211 "16"^^ . _:b437200859 . _:b437201685 "2"^^ . _:b437200212 "16"^^ . _:b437200858 . _:b437201684 "2"^^ . _:b437200213 "16"^^ . _:b437200869 . . . _:b437201687 "2"^^ . . _:b437200214 "16"^^ . _:b437200868 . . _:b437200215 "15"^^ . _:b437201686 "2"^^ . _:b437200871 . _:b437201227 . _:b437200216 "15"^^ . _:b437201681 "2"^^ . . _:b437200870 . . _:b437201026 . _:b437200217 "15"^^ . _:b437201680 "2"^^ . _:b437200865 . . _:b437200689 . _:b437200384 . _:b437200218 "15"^^ . _:b437201683 "2"^^ . . _:b437201181 . _:b437200864 . _:b437201913 . _:b437200219 "14"^^ . _:b437201260 . _:b437201682 "2"^^ . _:b437200867 . _:b437200220 "14"^^ . _:b437201693 "2"^^ . _:b437200866 . . _:b437200221 "14"^^ . _:b437201562 . . _:b437201692 "2"^^ . . _:b437200877 . _:b437200222 "14"^^ . . _:b437201695 "2"^^ . _:b437200779 . _:b437200876 . _:b437200223 "14"^^ . _:b437201694 "2"^^ . _:b437200879 . _:b437200224 "14"^^ . . _:b437201689 "2"^^ . _:b437200878 . _:b437200231 "13"^^ . _:b437200698 . _:b437201688 "2"^^ . _:b437200873 . _:b437200230 "13"^^ . . _:b437201691 "2"^^ . . _:b437200872 . _:b437200229 "13"^^ . _:b437201690 "2"^^ . _:b437200875 . _:b437201688 . _:b437200228 "13"^^ . _:b437201190 . _:b437201701 "2"^^ . _:b437200874 . _:b437200227 "13"^^ . . _:b437201700 "2"^^ . _:b437200885 . _:b437200226 "13"^^ . _:b437201703 "2"^^ . _:b437200884 . . _:b437200225 "13"^^ . _:b437200744 . _:b437201702 "2"^^ . _:b437201378 . _:b437200887 . . _:b437201705 . . _:b437201697 "2"^^ . _:b437200239 "12"^^ . _:b437200886 . _:b437201696 "2"^^ . _:b437200238 "12"^^ . _:b437200881 . . _:b437201699 "2"^^ . _:b437201770 . _:b437200237 "12"^^ . _:b437200880 . . _:b437201698 "2"^^ . _:b437200883 . _:b437200236 "13"^^ . . . _:b437201709 "2"^^ . _:b437200882 . _:b437200235 "13"^^ . _:b437201389 . _:b437201708 "2"^^ . _:b437200893 . _:b437200234 "13"^^ . _:b437201711 "2"^^ . _:b437200892 . _:b437200233 "13"^^ . _:b437201710 "2"^^ . _:b437200895 . _:b437200232 "13"^^ . _:b8124700 . _:b437201356 . _:b437201705 "2"^^ . _:b437201369 . _:b437200247 "12"^^ . _:b437201096 . _:b437200894 . _:b437201704 "2"^^ . _:b437200246 "12"^^ . _:b437200889 . . _:b437201707 "2"^^ . _:b437200245 "12"^^ . _:b437200888 . _:b437201706 "2"^^ . _:b437200244 "12"^^ . _:b437200891 . _:b437201164 . _:b437201311 . _:b437201717 "2"^^ . _:b437200243 "12"^^ . _:b437200890 . _:b437200658 . _:b437201716 "2"^^ . _:b437200242 "12"^^ . . _:b437200389 . . . _:b437201719 "2"^^ . _:b437200241 "12"^^ . _:b437200550 . _:b437200388 . _:b437201718 "2"^^ . _:b437200240 "12"^^ . _:b437200391 . _:b437200255 "11"^^ . _:b437201713 "2"^^ . _:b437200390 . _:b437200254 "11"^^ . _:b437201712 "2"^^ . _:b437200385 . . _:b437200253 "11"^^ . _:b437201715 "2"^^ . . _:b437200384 . _:b437200648 . _:b437200252 "11"^^ . _:b437201714 "2"^^ . _:b437200387 . _:b437200251 "11"^^ . _:b437201725 "2"^^ . . _:b437200386 . _:b437201465 . _:b437200250 "11"^^ . _:b8124644 "The absence of any other significant phenotype associated with a lack of CCR5 (>>5<<\u20137) has spurred the development of therapies aimed at blocking the virus-CCR5 interaction, and CCR5 antagonists have proved to be an effective salvage therapy in patients with drug-resistant strains of HIV-1 (8)." . _:b437201724 "2"^^ . _:b437200397 . _:b437201755 . _:b437200249 "11"^^ . _:b437201727 "2"^^ . _:b437200396 . . _:b437201726 "2"^^ . _:b437200343 . _:b437200399 . . _:b437200248 "12"^^ . _:b437200263 "10"^^ . _:b437201247 . _:b437201587 . _:b437201721 "2"^^ . _:b437200398 . _:b437201939 . _:b437200262 "11"^^ . _:b437201720 "2"^^ . _:b437201385 . _:b437200393 . _:b437200261 "11"^^ . _:b437201723 "2"^^ . _:b437200392 . _:b437200260 "11"^^ . _:b437201722 "2"^^ . _:b437200395 . _:b437200832 . _:b437200259 "11"^^ . _:b437201733 "2"^^ . _:b437200394 . . _:b437200258 "11"^^ . _:b437201732 "2"^^ . _:b437200405 . _:b437201573 . _:b437200257 "11"^^ . _:b437201735 "2"^^ . _:b437200404 . _:b437200256 "11"^^ . _:b437201734 "2"^^ . _:b437200407 . _:b437200271 "10"^^ . . _:b437201729 "2"^^ . _:b437200406 . _:b437201712 . _:b437200270 "10"^^ . _:b437201728 "2"^^ . _:b437200401 . _:b437200801 . _:b437200292 . _:b437200269 "10"^^ . . _:b437201731 "2"^^ . _:b437200400 . _:b437200794 . _:b437200268 "10"^^ . _:b437201502 . . _:b437201730 "2"^^ . . _:b437200403 . _:b437200267 "10"^^ . _:b437200539 . . . _:b437201741 "2"^^ . _:b437200402 . _:b437200266 "10"^^ . _:b437201740 "2"^^ . _:b437200413 . _:b437201493 . _:b437200497 . _:b437200265 "10"^^ . _:b437201743 "2"^^ . _:b437200412 . _:b437200890 . _:b437200264 "10"^^ . _:b437200860 . _:b437201742 "2"^^ . _:b437200415 . _:b437200279 "10"^^ . _:b437201737 "2"^^ . _:b437200414 . _:b437200278 "10"^^ . _:b437201736 "2"^^ . _:b437200409 . _:b437200277 "10"^^ . _:b437201739 "2"^^ . _:b437201844 . _:b437200408 . . _:b437200276 "10"^^ . _:b437201539 . _:b437201738 "2"^^ . _:b437200411 . . _:b437200275 "10"^^ . _:b437201749 "2"^^ . . _:b437200410 . . _:b437200274 "10"^^ . _:b437201309 . _:b437201748 "2"^^ . . _:b437200421 . _:b437200273 "10"^^ . _:b437201751 "2"^^ . _:b8124660 "Since DSB formation requires only transient expression of ZFNs, gene delivery methods that are suitable to express ZFNs include plasmid DNA nucleofection, integrase-defective lentiviral vectors and adenoviral vectors (16,>>19<<,21). Although non-viral methods such as nucleofection are attractive, this procedure has previously been associated with relatively high rates of toxicity in human CD34+ HSC and loss of engraftment potential (22), although more recently," . _:b437200420 . _:b8124644 . _:b437200272 "10"^^ . _:b437201750 "2"^^ . _:b437200423 . _:b8124645 . . . _:b437201745 "2"^^ . _:b437200422 . _:b8124646 . _:b437200286 "9"^^ . _:b437201744 "2"^^ . _:b437200417 . _:b8124647 . _:b437200287 "9"^^ . _:b437201747 "2"^^ . _:b437200416 . . _:b437200284 "9"^^ . _:b437201918 . _:b437201746 "2"^^ . . _:b437200419 . _:b437200285 "9"^^ . _:b8124641 . _:b437201757 "2"^^ . _:b437200418 . _:b8124642 . _:b437200282 "9"^^ . _:b437201577 . _:b437201756 "2"^^ . _:b437200392 . _:b8124643 . _:b437200429 . _:b437200283 "9"^^ . . . _:b437201759 "2"^^ . _:b437201127 . _:b437200428 . _:b8124652 . _:b437200280 "9"^^ . _:b437201758 "2"^^ . _:b437200431 . _:b8124653 . . _:b437200761 . . _:b437200281 "9"^^ . . _:b437200871 . _:b437201753 "2"^^ . _:b437200430 . _:b8124654 . _:b437200294 "9"^^ . _:b437201752 "2"^^ . _:b437200428 . _:b8124655 . _:b437201011 . _:b437200425 . _:b437200295 "9"^^ . . _:b437201755 "2"^^ . _:b437200424 . _:b8124648 . _:b437200844 . _:b437200292 "9"^^ . _:b437201669 . _:b8124714 . _:b437201754 "2"^^ . . _:b8124649 . _:b437200427 . _:b437200293 "9"^^ . _:b437201853 . _:b437201765 "2"^^ . _:b437200426 . _:b8124650 . _:b437200290 "9"^^ . _:b437200908 . _:b437201764 "2"^^ . _:b437200437 . _:b8124651 . _:b437200291 "9"^^ . _:b437201156 . _:b437201767 "2"^^ . _:b437200436 . _:b8124660 . _:b437200288 "9"^^ . _:b437200572 . _:b437201766 "2"^^ . _:b437200439 . _:b8124661 . _:b437201306 . _:b437200565 . _:b437200289 "9"^^ . _:b437200387 . _:b437201761 "2"^^ . _:b437200302 "8"^^ . _:b8124662 . _:b437200438 . _:b437200711 . _:b437201760 "2"^^ . _:b437200303 "8"^^ . _:b8124663 . . _:b437200433 . . . _:b437201763 "2"^^ . _:b437200300 "8"^^ . _:b8124656 . . _:b437200432 . _:b437201762 "2"^^ . _:b437200301 "8"^^ . _:b8124657 . _:b437200435 . . _:b437201773 "2"^^ . _:b437200298 "8"^^ . . _:b437200434 . _:b8124658 . . _:b437201772 "2"^^ . _:b437200299 "8"^^ . _:b8124659 . _:b437200445 . _:b437201652 . _:b437201775 "2"^^ . _:b8124666 "Similar results were also achieved using CD34+ cells isolated from fetal liver (data not shown). Previous studies in both cell lines (16) and primary human T cells (>>19<<) have shown that the percentage of bi-allelically modified cells in a population treated with ZFNs is between 30\u201340% of the total number of disrupted alleles detected by the Cel I assay, so that we estimate that 5\u20137% of the HSC would be" . _:b437200444 . _:b8124668 . . _:b437200296 "9"^^ . _:b437201774 "2"^^ . _:b437200447 . _:b8124669 . _:b437200297 "9"^^ . _:b437201319 . . _:b437201769 "2"^^ . _:b8124726 "It has been suggested that low level virus replication continues in certain sanctuary sites, even in well-controlled patients on ART (43,>>68<<), and this could provide a low level of selection, although drug intensification trials have not provided evidence of any such ongoing HIV-1 replication (69)." . _:b437200310 "8"^^ . _:b8124670 . . _:b437200446 . . _:b437201768 "2"^^ . _:b437200311 "8"^^ . _:b8124671 . _:b437200441 . _:b437201243 . . _:b437201636 . _:b437201771 "2"^^ . _:b437200308 "8"^^ . _:b8124664 . _:b437200440 . . _:b437201770 "2"^^ . _:b437200309 "8"^^ . _:b8124665 . _:b437200443 . _:b437201781 "2"^^ . _:b437200306 "8"^^ . _:b8124666 . _:b437200442 . _:b437201780 "2"^^ . _:b437200307 "8"^^ . _:b8124667 . _:b437200453 . . _:b437201783 "2"^^ . _:b437201402 . _:b437200304 "8"^^ . _:b437200452 . _:b437201141 . _:b8124663 "are attractive, this procedure has previously been associated with relatively high rates of toxicity in human CD34+ HSC and loss of engraftment potential (22), although more recently, less toxic outcomes have been described (>>23<<\u201325). We evaluated different parameters in order to identify nucleofection conditions that allowed the efficient disruption of the CCR5 gene by ZFNs, while limiting the associated toxicity." . _:b437201782 "2"^^ . _:b437200305 "8"^^ . _:b437200455 . _:b437201113 . _:b437201777 "2"^^ . _:b437200318 "8"^^ . _:b437200454 . _:b437201776 "2"^^ . _:b437200319 "8"^^ . _:b437200449 . _:b437201925 . . _:b437200800 . _:b437201779 "2"^^ . _:b437200316 "8"^^ . _:b437201950 . _:b437200448 . _:b437201924 . _:b437201778 "2"^^ . _:b437200317 "8"^^ . _:b8124705 "By using an optimized nucleofection procedure, we were able to overcome the technical challenges to ZFN-induced genome editing in HSC that have previously been reported (>>21<<) and achieve, on average, disruption at 17% of the loci." . _:b437200451 . _:b437201733 . _:b437201927 . _:b437201789 "2"^^ . _:b437200314 "8"^^ . _:b437200450 . _:b437201926 . _:b437201788 "2"^^ . _:b437200315 "8"^^ . _:b437200461 . _:b437201921 . _:b437201791 "2"^^ . _:b437200312 "8"^^ . _:b437200460 . _:b437201920 . _:b437201790 "2"^^ . _:b437200313 "8"^^ . _:b437200463 . _:b437201923 . _:b437201785 "2"^^ . _:b437200326 "8"^^ . _:b437200462 . _:b437201922 . . _:b437201017 . _:b437200355 . _:b437201784 "2"^^ . _:b437200699 . _:b437200327 "8"^^ . _:b437200457 . _:b437201933 . _:b437201787 "2"^^ . _:b437200324 "8"^^ . _:b437200456 . _:b437201932 . _:b437201786 "2"^^ . _:b437200325 "8"^^ . _:b437200459 . _:b437201935 . . _:b437201797 "2"^^ . _:b437200322 "8"^^ . _:b437200458 . _:b437200480 . _:b437201934 . . _:b437201796 "2"^^ . _:b437200323 "8"^^ . _:b437200469 . . . _:b8124683 "As previously reported for this ZFN pair (>>19<<), a high proportion (13 out of 59) of the mutated loci contained a characteristic 5bp duplication, with the remaining 46 clones comprising 36 unique sequences." . _:b437201929 . . _:b437201799 "2"^^ . _:b437201698 . _:b437200320 "8"^^ . _:b437200468 . _:b437201928 . . _:b437201798 "2"^^ . _:b437201461 . . _:b8124687 . _:b437200471 . _:b437200321 "8"^^ . . _:b437201931 . . _:b437200334 "7"^^ . _:b437201793 "2"^^ . _:b437200470 . _:b437201930 . _:b8124719 "However only a handful of cases of HIV-1 infection of CCR5\u039432 homozygotes have been reported (>>63<<,64). CXCR4-tropic viruses have been associated with accelerated disease progression (65), so that selection for such strains could be an undesirable consequence of targeting CCR5. However, this outcome is not generally observed in" . . _:b437201792 "2"^^ . _:b437200335 "7"^^ . _:b437200465 . _:b437201941 . . _:b437200481 . _:b437201795 "2"^^ . _:b437200332 "8"^^ . _:b437200464 . _:b437201940 . . _:b437201794 "2"^^ . _:b437200333 "8"^^ . _:b437200467 . _:b437200864 . _:b437201943 . _:b437200804 . _:b437201805 "2"^^ . _:b437201610 . _:b437200330 "8"^^ . _:b437201103 . _:b437200466 . _:b437201942 . _:b437201804 "2"^^ . _:b437200331 "8"^^ . _:b437200477 . _:b437201937 . . _:b437201807 "2"^^ . _:b437200328 "8"^^ . _:b437200476 . _:b437201936 . _:b437201806 "2"^^ . _:b437200329 "8"^^ . _:b437200969 . _:b437200479 . _:b437201939 . _:b437200342 "7"^^ . _:b437201801 "2"^^ . _:b437200478 . _:b437201938 . _:b437200343 "7"^^ . _:b437201800 "2"^^ . _:b437200473 . _:b437201949 . . _:b437200340 "7"^^ . _:b437201803 "2"^^ . _:b437200472 . _:b437201948 . . _:b437200341 "7"^^ . _:b437201802 "2"^^ . _:b437200475 . _:b437201951 . . _:b437200338 "7"^^ . _:b437201813 "2"^^ . _:b437200474 . . _:b437201950 . _:b437200339 "7"^^ . _:b437201812 "2"^^ . _:b437200526 . _:b437200485 . _:b437201945 . . _:b437200336 "7"^^ . _:b437201815 "2"^^ . _:b437200484 . _:b437201944 . . _:b437200337 "7"^^ . _:b437201814 "2"^^ . _:b437200912 . _:b437200487 . _:b437201947 . _:b437200350 "7"^^ . _:b437201809 "2"^^ . . _:b437200486 . _:b437201946 . _:b437201545 . _:b437200351 "7"^^ . _:b437201808 "2"^^ . _:b437200481 . . _:b437201957 . _:b437200348 "7"^^ . _:b437201811 "2"^^ . _:b437200480 . _:b437200464 . _:b437201956 . _:b437200349 "7"^^ . _:b437201810 "2"^^ . _:b437200483 . _:b437200996 . . _:b437201959 . . _:b437200346 "7"^^ . _:b437201821 "2"^^ . . _:b437200482 . _:b437201958 . _:b437200347 "7"^^ . _:b437201820 "2"^^ . _:b437200493 . _:b437201953 . _:b437200344 "7"^^ . _:b437201823 "2"^^ . . _:b437200492 . _:b437201952 . _:b437200345 "7"^^ . _:b437201822 "2"^^ . _:b437200495 . . _:b437201955 . _:b437200358 "7"^^ . _:b437201817 "2"^^ . _:b437200494 . . _:b437201954 . . _:b437201816 "2"^^ . _:b437200359 "7"^^ . . _:b437200489 . _:b437201965 . . _:b437201816 . _:b437200356 "7"^^ . _:b437201819 "2"^^ . _:b437200488 . _:b8124722 . _:b437201964 . _:b437200357 "7"^^ . _:b437201818 "2"^^ . _:b437200491 . _:b437201967 . _:b437200354 "7"^^ . _:b437201829 "2"^^ . _:b437200490 . _:b437201028 . _:b437200950 . _:b437201966 . _:b437200355 "7"^^ . _:b437201828 "2"^^ . _:b437200501 . _:b437201701 . _:b437200535 . . _:b437201961 . _:b437200352 "7"^^ . _:b437201831 "2"^^ . . . _:b437200500 . _:b437201775 . _:b437201960 . _:b437200353 "7"^^ . _:b437201830 "2"^^ . _:b437200503 . _:b437201963 . . . _:b437201825 "2"^^ . _:b437201608 . _:b437200366 "7"^^ . _:b437200502 . _:b437201337 . _:b437201962 . _:b437200367 "6"^^ . _:b437201824 "2"^^ . _:b437200497 . _:b437201973 . _:b437200914 . _:b437200364 "7"^^ . _:b437201827 "2"^^ . . _:b437200496 . _:b437201972 . _:b437200364 . _:b437200365 "7"^^ . _:b437201826 "2"^^ . _:b437201515 . _:b437200382 . _:b437200499 . _:b437201975 . _:b437200362 "7"^^ . _:b437201837 "2"^^ . _:b437200498 . . . _:b437201974 . . _:b437200710 . _:b437200363 "7"^^ . _:b437201836 "2"^^ . _:b437200509 . _:b437201696 . _:b437201969 . . _:b437200360 "7"^^ . _:b437201839 "2"^^ . _:b437200508 . _:b437201968 . _:b437200361 "7"^^ . _:b437201838 "2"^^ . _:b437200511 . . _:b437201971 . _:b437200374 "6"^^ . _:b437201833 "2"^^ . . _:b437200510 . _:b437201970 . _:b437200375 "6"^^ . _:b437200799 . _:b437201832 "2"^^ . _:b437201162 . _:b437200505 . . _:b437201981 . _:b437200372 "6"^^ . . _:b437201835 "2"^^ . _:b437200504 . . _:b437201980 . _:b437200373 "6"^^ . . . _:b437201834 "2"^^ . . _:b8124648 . _:b437200507 . . _:b437201983 . _:b437200370 "6"^^ . . _:b437201845 "2"^^ . . _:b437200506 . . _:b437201982 . . _:b437200371 "6"^^ . _:b437201844 "2"^^ . _:b437200517 . _:b437201977 . _:b437200368 "6"^^ . _:b437201847 "2"^^ . _:b437200516 . _:b437200613 . _:b437201976 . _:b437200369 "6"^^ . _:b437201846 "2"^^ . _:b8124710 . _:b437200519 . _:b437201979 . _:b437200382 "6"^^ . _:b8124703 "ZFNs can be designed to bind to a specific genomic DNA sequence and, by generating a targeted DSB, can affect the efficient, specific and permanent knockout of the targeted gene (19,>>45<<\u201347). In contrast to other genetic approaches to the control of HIV-1, ZFN-mediated gene disruption is a one-time, \u201Chit-and-run\u201D process that requires only transient expression of the ZFNs in HSC during a brief period of ex vivo culture." . _:b437201841 "2"^^ . _:b437200518 . _:b437201978 . _:b437200383 "6"^^ . . _:b437201840 "2"^^ . _:b437200513 . _:b437200933 . . _:b437201989 . _:b437200380 "6"^^ . _:b437201843 "2"^^ . _:b437201985 . _:b437200512 . _:b437200900 . _:b437201988 . _:b437201647 . _:b437200381 "6"^^ . _:b437201560 . _:b437201842 "2"^^ . _:b437200515 . . _:b437201991 . _:b437200378 "6"^^ . _:b437201853 "2"^^ . _:b437200514 . _:b8124670 . _:b437201759 . _:b437201990 . _:b437200379 "6"^^ . _:b437201228 . _:b437200521 . _:b437201852 "2"^^ . _:b437200525 . _:b437201985 . _:b437200376 "6"^^ . _:b437200478 . _:b437201855 "2"^^ . . _:b437200524 . _:b437201984 . _:b437200377 "6"^^ . _:b437201854 "2"^^ . _:b437200527 . _:b437201407 . _:b437201987 . . _:b437201849 "2"^^ . _:b437200526 . _:b437201986 . _:b437201848 "2"^^ . _:b437200521 . . _:b437201997 . . . _:b437201851 "2"^^ . _:b437200520 . _:b437201996 . _:b437200884 . _:b437200690 . _:b437201061 . _:b437201850 "2"^^ . _:b437201342 . _:b437200523 . _:b437200374 . _:b437201999 . _:b437201292 . _:b437201861 "2"^^ . _:b437200522 . . _:b437201998 . . _:b437201860 "2"^^ . _:b437200380 . _:b437200446 . _:b437200533 . . _:b437201993 . . _:b437201863 "2"^^ . _:b437200532 . _:b437201992 . _:b437201862 "2"^^ . _:b437200535 . _:b437201995 . _:b437200627 . _:b437201151 . . _:b437201857 "2"^^ . _:b437200534 . _:b437201994 . _:b437201856 "2"^^ . _:b437200529 . _:b437202005 . . _:b437201859 "2"^^ . _:b437200528 . _:b437201192 . _:b437202004 . _:b437201858 "2"^^ . _:b437200452 . _:b437200531 . _:b437201675 . _:b437202007 . _:b437201906 . . _:b437200654 . _:b437201869 "2"^^ . _:b437200530 . _:b437202006 . _:b437201868 "2"^^ . _:b437200541 . _:b437200897 . _:b437202001 . . _:b437201871 "2"^^ . _:b437200540 . . _:b437202000 . . . _:b437201870 "2"^^ . _:b437200543 . . _:b437202003 . _:b437201865 "2"^^ . . . _:b437200542 . _:b437201404 . _:b437202002 . _:b437201864 "2"^^ . _:b437200537 . . _:b437201867 "2"^^ . . _:b437200536 . _:b437201866 "2"^^ . _:b437200539 . _:b437201877 "2"^^ . _:b437200538 . . . _:b437201876 "2"^^ . _:b437200549 . _:b437201680 . _:b437201879 "2"^^ . _:b437200548 . _:b437202008 . _:b437201878 "2"^^ . _:b437200551 . _:b437200425 . _:b437200681 . _:b437201873 "2"^^ . _:b437200550 . _:b437201872 "2"^^ . _:b437200545 . . _:b437201875 "2"^^ . _:b437200544 . . _:b437201874 "2"^^ . _:b437201526 . _:b437200547 . . . _:b437201885 "2"^^ . _:b437200546 . . _:b437201884 "2"^^ . _:b437200557 . _:b437201887 "2"^^ . _:b437200556 . _:b8124658 "results" . _:b437201173 . . _:b437201886 "2"^^ . _:b437200559 . . . . _:b437201881 "2"^^ . _:b437201055 . _:b437200558 . . . _:b437201880 "2"^^ . _:b437200553 . _:b437201883 "2"^^ . _:b437200552 . . . . _:b437201882 "2"^^ . _:b437200555 . . _:b437201893 "2"^^ . _:b437200554 . _:b437201892 "2"^^ . _:b437200565 . . _:b437201895 "2"^^ . _:b437200564 . _:b437201109 . . _:b437200463 . . _:b437201894 "2"^^ . _:b437200567 . _:b437201425 . . _:b437201075 . _:b437201889 "2"^^ . _:b437200566 . . . _:b437201888 "2"^^ . _:b437200561 . . _:b437201891 "2"^^ . _:b437201459 . _:b437200560 . _:b437201890 "2"^^ . _:b437200563 . _:b437200998 . _:b437201901 "2"^^ . _:b437200562 . . _:b437201900 "2"^^ . _:b437200573 . . . _:b437201903 "2"^^ . _:b437200572 . _:b437201423 . _:b437201902 "2"^^ . _:b437200575 . . _:b437201897 "2"^^ . _:b437200574 . . . _:b437201896 "2"^^ . _:b437201308 . _:b437200569 . _:b437201661 . _:b437201208 . _:b437201899 "2"^^ . _:b437200568 . _:b437200390 . _:b437201898 "2"^^ . _:b437200571 . _:b437201122 . _:b437201909 "2"^^ . _:b437200570 . _:b437201908 "2"^^ . _:b437200581 . _:b437201317 . _:b437201877 . . _:b437201911 "2"^^ . _:b437200580 . _:b437201150 . _:b437201751 . . _:b437201910 "2"^^ . _:b437200583 . . . _:b437201905 "2"^^ . _:b437200582 . _:b437200490 . _:b437200661 . . . _:b437201904 "2"^^ . . _:b437200577 . _:b437201907 "2"^^ . _:b437200576 . _:b437201906 "2"^^ . _:b437200579 . _:b437200254 . . _:b437201917 "2"^^ . . _:b437200578 . _:b437200733 . _:b437200899 . _:b437201916 "2"^^ . . _:b437200589 . . _:b437201919 "2"^^ . _:b437200588 . _:b437200717 . _:b437201918 "2"^^ . _:b437200211 . _:b437200591 . . _:b437200278 . _:b437201913 "2"^^ . _:b437200590 . _:b437201912 "2"^^ . _:b437200585 . _:b437201915 "2"^^ . . _:b437200584 . _:b437201803 . . _:b437201914 "2"^^ . _:b437200587 . _:b437201354 . _:b437200901 "3"^^ . _:b437200586 . . _:b437200796 . _:b437200597 . _:b437200900 "3"^^ . _:b437200762 . _:b437201146 . _:b437200903 "3"^^ . _:b437200596 . . _:b437200902 "3"^^ . _:b437200599 . _:b437200897 "3"^^ . _:b437200598 . _:b437200776 . _:b437201323 . _:b437200896 "3"^^ . _:b437201646 . _:b437200593 . . . _:b437200702 . _:b437200619 . _:b437200899 "3"^^ . _:b437200592 . _:b437201834 . _:b437200898 "3"^^ . . _:b437200595 . _:b437200378 . _:b437200909 "3"^^ . _:b437200594 . _:b437200908 "3"^^ . _:b437200605 . _:b437200295 . _:b437201397 . _:b437200420 . . _:b437200604 . _:b437200911 "3"^^ . _:b437201543 . _:b437200773 . _:b437200910 "3"^^ . _:b437200607 . _:b437200905 "3"^^ . _:b437201697 . _:b437200606 . . _:b437200904 "3"^^ . _:b437200601 . . _:b437200907 "3"^^ . _:b437200600 . _:b437201153 . _:b437200906 "3"^^ . . _:b437200603 . _:b437201364 . _:b8124656 "approach, we optimized the delivery of CCR5-specific ZFNs to human HSC and analyzed the consequences of this modification for the stem cells by transplantation into NOD/SCID/IL2r\u03B3null (NSG) mice, which support both human hematopoiesis (>>20<<) and HIV-1 infection (13)." . _:b437200917 "3"^^ . _:b437200602 . _:b437200916 "3"^^ . _:b8124707 "Mouse studies have not found any deleterious phenotype associated with loss of CCR2 (>>48<<), and human genetic studies have even suggested a beneficial phenotype from the loss of this gene in HIV-infected individuals (49)." . _:b437200613 . _:b437200314 . . _:b437200612 . _:b437200919 "3"^^ . . _:b437200918 "3"^^ . _:b437200615 . _:b437201358 . _:b437201078 . _:b437200913 "3"^^ . . _:b437200614 . _:b437200912 "3"^^ . _:b437201860 . _:b437200609 . _:b437201850 . . _:b437200915 "3"^^ . _:b437200608 . _:b437200216 . . _:b437201235 . _:b437201981 . . _:b437200611 . _:b437200914 "3"^^ . _:b437201638 . _:b437200818 . _:b437201424 . . _:b437200610 . _:b437200925 "3"^^ . _:b437200205 . . . _:b437200372 . _:b437200621 . _:b437200924 "3"^^ . . _:b437201810 . _:b437200927 "3"^^ . . _:b437200620 . . _:b437201797 . . _:b437200926 "3"^^ . . _:b437200623 . _:b437200437 . _:b437200775 . _:b437200989 . _:b437200921 "3"^^ . _:b437200622 . _:b437200617 . _:b437200920 "3"^^ . . _:b437200923 "3"^^ . _:b437200616 . . . _:b8124644 . _:b437200317 . _:b437200922 "3"^^ . _:b437200619 . _:b437200618 . _:b437200933 "3"^^ . . _:b437200629 . _:b437201994 . _:b437200932 "3"^^ . . _:b437200628 . _:b437200935 "3"^^ . . _:b437200934 "3"^^ . _:b437200631 . . _:b437200630 . _:b437200929 "3"^^ . _:b437201883 . _:b437200625 . _:b437200928 "3"^^ . _:b437200591 . _:b437200624 . _:b437200931 "3"^^ . . _:b437200803 . _:b437200930 "3"^^ . _:b437200627 . . _:b437201841 . _:b437200626 . _:b437200941 "3"^^ . . _:b437200637 . _:b437200940 "3"^^ . . . _:b437200943 "3"^^ . _:b437200636 . _:b437200639 . _:b437201645 . _:b437200942 "3"^^ . . _:b437200638 . _:b437200937 "3"^^ . _:b437201525 . _:b437200633 . _:b437200936 "3"^^ . _:b437201598 . _:b437200877 . _:b437200632 . _:b437200939 "3"^^ . _:b437200635 . _:b437200938 "3"^^ . _:b437200917 . _:b437201485 . _:b437200949 "3"^^ . _:b437200634 . _:b437200742 . _:b437200948 "3"^^ . _:b8124663 . _:b437200951 "3"^^ . _:b437200950 "3"^^ . . _:b437200945 "3"^^ . _:b437201615 . _:b437200944 "3"^^ . . _:b437201432 . _:b437200809 . _:b437200646 . . _:b437200947 "3"^^ . . _:b437200946 "3"^^ . _:b437200957 "3"^^ . . _:b437200956 "3"^^ . . . _:b437200959 "3"^^ . . _:b437200730 . _:b437201266 . _:b437200958 "3"^^ . . . _:b437200953 "3"^^ . _:b437200952 "3"^^ . _:b8124690 "In addition, a common 5bp (pentamer) duplication that occurs following NHEJ repair of the ZFN-cleaved CCR5 locus (>>19<<) was detected by PCR. The first round PCR product generated during Cel I analysis was diluted 1:" . . . _:b437200955 "3"^^ . _:b437200954 "3"^^ . _:b437200965 "3"^^ . _:b437200892 . . _:b437200270 . _:b437200964 "3"^^ . . _:b437200967 "3"^^ . . . _:b437200966 "3"^^ . . _:b437200961 "3"^^ . . . _:b437201446 . _:b437200960 "3"^^ . . . _:b437200963 "3"^^ . _:b437201447 . . _:b437200962 "3"^^ . _:b437200973 "3"^^ . _:b437200972 "3"^^ . _:b437202000 . . _:b437200975 "3"^^ . . _:b437200974 "3"^^ . . _:b437200969 "3"^^ . . _:b437200754 . . _:b437200968 "3"^^ . . _:b437201599 . _:b437200971 "3"^^ . . _:b437200970 "3"^^ . . _:b437200713 . _:b437200981 "3"^^ . _:b437201420 . _:b437201792 . _:b437200980 "3"^^ . . _:b437200787 . _:b437200983 "3"^^ . _:b437200982 "3"^^ . . _:b437200977 "3"^^ . _:b437201305 . . . _:b437200976 "3"^^ . _:b437200520 . _:b437200338 . _:b437200979 "3"^^ . _:b8124656 . _:b437200940 . _:b437200978 "3"^^ . _:b437200959 . _:b437200929 . _:b437200791 . _:b437200989 "3"^^ . . _:b437201558 . _:b437201265 . _:b437200688 . _:b437201724 . . _:b437200952 . . . _:b437200988 "3"^^ . _:b437200199 . _:b437201884 . _:b8124669 . _:b437200991 "3"^^ . _:b437200394 . _:b437200990 "3"^^ . . _:b437200895 . . _:b437200635 . _:b437200985 "3"^^ . . . _:b437200984 "3"^^ . . _:b437200987 "3"^^ . _:b437200986 "3"^^ . _:b437201293 . _:b437200997 "3"^^ . _:b437200996 "3"^^ . _:b437200999 "3"^^ . _:b437201749 . _:b437200633 . _:b437200388 . _:b437200998 "3"^^ . . _:b437201927 . . _:b437200993 "3"^^ . _:b437200992 "3"^^ . . _:b437200995 "3"^^ . _:b437201331 . . _:b437200994 "3"^^ . . . _:b437201974 . _:b437201005 "3"^^ . . _:b437200506 . _:b437200189 . _:b437201004 "3"^^ . . . _:b437200188 . _:b437201007 "3"^^ . . _:b437201916 . _:b437201275 . _:b437201382 . _:b437200191 . _:b437201006 "3"^^ . _:b437201752 . _:b437200190 . _:b437201001 "3"^^ . _:b437201000 "3"^^ . . . _:b437201003 "3"^^ . _:b437201307 . _:b437200187 . _:b437201002 "3"^^ . _:b437200186 . _:b437201013 "3"^^ . _:b437200197 . _:b437201012 "3"^^ . . . _:b8124684 "virus in the blood, despite the acute loss of CD4+ cells, is reminiscent of observations in patients, were high viral load measurements in serum are typically observed during progression to AIDS, when T cell death is rapidly occurring (>>41<<). In contrast, CD4+ T cell levels rebounded in the ZFN-treated mice after the 2 week nadir, and recovered to normal levels by 4 weeks post-infection." . . _:b437201015 "3"^^ . _:b437200196 . _:b437200199 . _:b437201014 "3"^^ . _:b437200261 . . . _:b437201009 "3"^^ . _:b437200198 . _:b437201639 . _:b437200193 . _:b437201008 "3"^^ . _:b437200280 . _:b437200977 . _:b437201669 . . _:b437201011 "3"^^ . _:b437200192 . _:b437201668 . _:b437200195 . _:b437201010 "3"^^ . _:b437201671 . . . _:b437201021 "3"^^ . _:b437200194 . _:b437201670 . . _:b437200774 . _:b437201020 "3"^^ . _:b437200205 . . _:b437201665 . . _:b437200204 . _:b437201023 "3"^^ . _:b437201664 . _:b437200207 . _:b437201022 "3"^^ . _:b437200904 . _:b437201667 . . _:b437200206 . _:b437201017 "3"^^ . _:b437201666 . . _:b437200201 . _:b437201016 "3"^^ . _:b437201677 . _:b437200200 . _:b437201019 "3"^^ . _:b437201676 . _:b437200429 . _:b437201018 "3"^^ . _:b437200203 . _:b437201679 . _:b437200291 . . _:b437201029 "3"^^ . _:b437200202 . _:b437201678 . _:b437200831 . _:b437201028 "3"^^ . _:b437200213 . _:b437201673 . _:b437201773 . _:b437200212 . _:b437201067 . _:b437201031 "3"^^ . _:b437201672 . _:b437200869 . _:b437200238 . . _:b437201030 "3"^^ . _:b437200215 . _:b437201675 . _:b437200294 . _:b437200942 . _:b437200214 . _:b437201025 "3"^^ . _:b437201674 . _:b437200209 . _:b437201024 "3"^^ . _:b437201685 . _:b437200208 . _:b437201170 . _:b437201027 "3"^^ . . _:b437201684 . _:b437200211 . _:b437201026 "3"^^ . . _:b437201687 . _:b437200210 . _:b437201037 "3"^^ . _:b437201686 . _:b437200221 . _:b437201036 "3"^^ . . _:b437201681 . . _:b437201039 "3"^^ . _:b437200220 . _:b437201680 . _:b437200223 . _:b437201038 "3"^^ . _:b437201683 . _:b437201901 . _:b437200213 . _:b437200222 . _:b437201033 "3"^^ . _:b437201682 . _:b8124657 "delivery of CCR5-specific ZFNs to human HSC and analyzed the consequences of this modification for the stem cells by transplantation into NOD/SCID/IL2r\u03B3null (NSG) mice, which support both human hematopoiesis (20) and HIV-1 infection (>>13<<). Following challenge of the mice with a CCR5-tropic strain of HIV-1, we observed rapid selection for human cells that were CCR5-negative, a significant reduction in viral load and protection of human T cell populations in the key tissues" . _:b437200451 . _:b437201032 "3"^^ . _:b437200217 . _:b437201693 . _:b437200216 . _:b437201035 "3"^^ . _:b437201692 . _:b437201915 . _:b437200219 . _:b437201034 "3"^^ . . _:b437201695 . . _:b437201045 "3"^^ . _:b437200218 . . _:b437201694 . _:b437200229 . _:b437201044 "3"^^ . _:b437200823 . _:b437201689 . . _:b437200228 . _:b437201047 "3"^^ . _:b437201688 . _:b437200231 . _:b437201046 "3"^^ . _:b437201691 . _:b437200230 . _:b437201041 "3"^^ . _:b437201690 . _:b437200671 . _:b437201040 "3"^^ . _:b437200225 . _:b437201261 . _:b437201701 . _:b437200224 . _:b437201043 "3"^^ . _:b437200923 . _:b437201700 . . . _:b437200227 . _:b437201042 "3"^^ . _:b437201703 . . _:b437201575 . _:b437200672 . _:b437201053 "3"^^ . _:b437200226 . . _:b437201702 . . _:b437200548 . _:b437201052 "3"^^ . _:b437200237 . . _:b437201697 . . _:b437200236 . _:b437201500 . _:b437201055 "3"^^ . _:b437201696 . _:b437200200 . _:b437201054 "3"^^ . _:b437200239 . _:b437201699 . . . _:b437200238 . _:b437201049 "3"^^ . _:b437200449 . _:b437201698 . . _:b437200325 . _:b437200233 . _:b437201048 "3"^^ . _:b437201213 . _:b437201709 . _:b437200232 . _:b437201051 "3"^^ . _:b437201708 . . _:b437200235 . _:b437201050 "3"^^ . _:b437201711 . . _:b437201154 . . _:b437201061 "3"^^ . _:b437200234 . _:b437201710 . _:b437200245 . _:b437201060 "3"^^ . _:b437201705 . _:b437200244 . _:b437201063 "3"^^ . _:b437201704 . _:b437200247 . _:b8124680 . _:b437201062 "3"^^ . _:b437201707 . _:b437200246 . _:b437201057 "3"^^ . _:b437201706 . . . _:b437201056 "3"^^ . _:b437200241 . _:b437201717 . _:b437201799 . _:b437200240 . _:b437201059 "3"^^ . _:b437201716 . _:b437201851 . _:b437200253 . _:b8124676 . _:b437201058 "3"^^ . _:b437200243 . _:b437201719 . _:b437201391 . _:b437200242 . _:b437201640 . _:b437201069 "3"^^ . _:b437201718 . "PMC0" . _:b437200253 . _:b437201068 "3"^^ . _:b437201713 . . . _:b437201071 "3"^^ . _:b437200252 . _:b437201712 . _:b437200255 . _:b437201070 "3"^^ . _:b437201715 . _:b437201900 . _:b437200254 . _:b437201065 "3"^^ . _:b437201714 . . _:b437200413 . _:b437200592 . _:b437201064 "3"^^ . _:b437200249 . _:b437201725 . _:b437201350 . _:b437200578 . _:b437201067 "3"^^ . _:b437200248 . _:b437201724 . _:b437200251 . _:b437201443 . _:b437201066 "3"^^ . _:b437201727 . . _:b437200250 . _:b437201077 "3"^^ . . _:b437201726 . _:b437200261 . _:b437201076 "3"^^ . _:b437201721 . _:b437200260 . _:b437201079 "3"^^ . _:b437201720 . . _:b437200263 . _:b437201078 "3"^^ . _:b437201723 . _:b437200262 . _:b437201073 "3"^^ . _:b437201374 . _:b437201722 . . _:b437200257 . _:b437201216 . _:b437201072 "3"^^ . . _:b437201733 . _:b437200256 . _:b437201075 "3"^^ . . _:b437201732 . _:b437201223 . _:b437200259 . _:b437201074 "3"^^ . _:b437201735 . . _:b437201085 "2"^^ . _:b437200258 . _:b437201274 . _:b437201734 . _:b437201084 "2"^^ . _:b437200269 . . _:b437201729 . . _:b437201087 "2"^^ . _:b437200268 . _:b437201023 . _:b437201728 . _:b437201086 "2"^^ . _:b437200271 . _:b437201731 . . _:b437200270 . _:b437201081 "3"^^ . . _:b437201730 . _:b437200265 . _:b437201080 "3"^^ . _:b437201741 . . _:b437201083 "2"^^ . _:b437200264 . . _:b437201740 . _:b437201082 "2"^^ . . _:b437200267 . _:b437201743 . _:b437201093 "2"^^ . . _:b437200266 . _:b437201742 . _:b437201092 "2"^^ . _:b437200277 . _:b437201393 . _:b437201737 . _:b437201509 . _:b437201095 "2"^^ . _:b437201145 . _:b437200276 . _:b437201231 . _:b437201736 . _:b437201094 "2"^^ . _:b437200279 . _:b437201739 . _:b437201089 "2"^^ . _:b437200278 . _:b437201668 . _:b437201738 . _:b437201088 "2"^^ . _:b437200273 . . _:b437201749 . . _:b437201091 "2"^^ . . _:b437200272 . _:b437201703 . _:b437201748 . _:b437201090 "2"^^ . _:b437200913 . _:b437200275 . _:b437201751 . _:b437201101 "2"^^ . . _:b437200274 . . _:b437201750 . . _:b437201100 "2"^^ . . _:b8124682 "ZFN-induced DSBs repaired by the NHEJ pathway result in highly heterogeneous changes at the targeted locus (>>19<<). We used this property to investigate whether the CCR5-negative cells that developed in mice transplanted with ZFN-treated HSC were polyclonal in origin." . _:b437200285 . _:b437201745 . _:b437201574 . _:b437201103 "2"^^ . _:b437201201 . _:b437200284 . _:b437201744 . _:b437201102 "2"^^ . _:b437200287 . _:b437201747 . _:b437201097 "2"^^ . _:b437200286 . _:b437201746 . _:b437200792 . _:b437201527 . _:b437201096 "2"^^ . _:b437200281 . _:b437201757 . _:b437201099 "2"^^ . _:b437200280 . _:b437201756 . . . _:b437201098 "2"^^ . _:b437200283 . _:b437201759 . _:b437200721 . _:b437201109 "2"^^ . _:b437200282 . _:b437201758 . _:b437201108 "2"^^ . _:b437200293 . _:b8124698 "regimens that are not well tolerated by a significant percentage of patients (42), and even successful adherence to ART does not eradicate the virus, so that a rapid rebound in HIV-1 levels can occur if therapy is discontinued (>>43<<). Consequently, an alternate approach to controlling HIV-1 replication that is being considered is the engineering of the body\u2019s immune cells to be resistant to infection by the virus (44)." . _:b437201753 . . _:b437201111 "2"^^ . . _:b437200743 . _:b437200292 . . _:b437201752 . _:b437201592 . . . . _:b437201110 "2"^^ . _:b437200295 . _:b437201755 . _:b437201766 . _:b437201105 "2"^^ . _:b437200294 . _:b437200655 . _:b437201754 . _:b437201678 . _:b437200310 . _:b437201104 "2"^^ . . _:b437200289 . _:b437201765 . _:b437200951 . _:b437201107 "2"^^ . _:b437200288 . _:b437201522 . _:b437201764 . _:b437201106 "2"^^ . _:b437201999 . _:b437201895 . _:b437200291 . . _:b437201767 . . _:b437201117 "2"^^ . _:b437200290 . _:b437201204 . _:b437201766 . _:b437201437 . _:b437201116 "2"^^ . _:b437200301 . _:b437201761 . _:b437201119 "2"^^ . _:b437200300 . . _:b437201760 . _:b437201561 . . _:b437202006 . _:b437201118 "2"^^ . _:b437200303 . _:b437201322 . _:b437201763 . _:b437201837 . . _:b437201113 "2"^^ . _:b437200302 . _:b437201762 . _:b437201112 "2"^^ . _:b437200297 . _:b437201773 . _:b437201289 . _:b437201115 "2"^^ . _:b437200729 . _:b8124648 . _:b437200296 . . _:b437201772 . _:b437201114 "2"^^ . _:b437200299 . _:b437201775 . _:b437201125 "2"^^ . _:b437200298 . _:b437201774 . _:b437201175 . _:b437201124 "2"^^ . _:b437200309 . _:b437201769 . . _:b437201554 . _:b437201127 "2"^^ . _:b437200308 . . _:b437201768 . . _:b437201126 "2"^^ . _:b437201362 . _:b437200311 . _:b437201771 . _:b437201121 "2"^^ . _:b437201512 . _:b437200310 . _:b437201770 . _:b437201120 "2"^^ . _:b437200305 . _:b437201781 . . _:b437201123 "2"^^ . _:b437200304 . _:b437201602 . . _:b437201780 . _:b437201468 . _:b437201122 "2"^^ . _:b437200307 . . _:b437201783 . . _:b437200598 . _:b437201133 "2"^^ . _:b437200306 . _:b437201334 . _:b437200188 . _:b437201782 . _:b437201132 "2"^^ . _:b437200317 . . . _:b437201777 . _:b437200309 . _:b437201135 "2"^^ . _:b437200316 . _:b437201776 . _:b437200623 . . _:b437200599 . _:b437201817 . _:b437201134 "2"^^ . _:b437200319 . _:b437201779 . _:b437200346 . _:b437200435 . _:b437201129 "2"^^ . . _:b437200318 . _:b437201778 . . _:b437201128 "2"^^ . _:b437200313 . _:b437201789 . _:b437201131 "2"^^ . . _:b437200312 . _:b437200960 . _:b437201788 . . _:b437201130 "2"^^ . _:b437200315 . _:b437201791 . . . . _:b437201141 "2"^^ . _:b437200314 . _:b437201795 . . _:b437201790 . _:b437200381 . _:b437201140 "2"^^ . _:b437200325 . _:b437201785 . _:b437201143 "2"^^ . . _:b437201409 . _:b437200324 . _:b437201784 . _:b437201142 "2"^^ . _:b8124715 "Of note, certain HIV-infected patient populations, such as AIDS lymphoma patients, already undergo full ablation and autologous HSC rescue as part of their therapy (58), and thereby present an opportunity for HSC-based gene therapies (>>44<<). In addition, the experience of autologous HSC transplantation in gene therapy for patients with genetic diseases such as ADA-SCID (59\u201360), chronic granulomatous disease (61) and X-linked adrenoleukodystrophy (62), is that" . _:b437200564 . _:b437200327 . _:b437201787 . _:b437201137 "2"^^ . _:b437200300 . _:b437201142 . _:b437200326 . _:b437200919 . _:b437201786 . _:b437201136 "2"^^ . _:b437200321 . _:b437201797 . . . _:b437201139 "2"^^ . _:b437200320 . _:b437201796 . _:b8124671 . . _:b437201138 "2"^^ . _:b437200323 . . _:b437201799 . . _:b437201149 "2"^^ . _:b437200322 . _:b437201798 . _:b437201148 "2"^^ . _:b437200333 . _:b437201793 . _:b437200473 . . _:b437201151 "2"^^ . _:b437200332 . _:b437201301 . _:b437200573 . _:b437201792 . . _:b437201010 . . _:b437201150 "2"^^ . _:b437200335 . . _:b437201795 . _:b437200336 . _:b437201145 "2"^^ . _:b437200334 . _:b437201794 . . _:b437201144 "2"^^ . _:b437200329 . _:b437201805 . . . _:b437201147 "2"^^ . . _:b437200328 . _:b437201804 . . . _:b437201146 "2"^^ . _:b437200331 . . _:b437201807 . _:b437201157 "2"^^ . _:b437200330 . _:b437201806 . _:b437200686 . _:b437201156 "2"^^ . _:b437200341 . _:b437201801 . . _:b437201159 "2"^^ . _:b437200340 . _:b437201800 . _:b437200885 . _:b437201158 "2"^^ . _:b437200343 . _:b437201803 . _:b437201874 . _:b8124708 "Mouse studies have not found any deleterious phenotype associated with loss of CCR2 (48), and human genetic studies have even suggested a beneficial phenotype from the loss of this gene in HIV-infected individuals (>>49<<). Although not specifically analyzed in our study, it is likely that the use of these same CCR5 ZFN reagents in HSC will result in similar, low levels of off-target events, the consequences of which will need to be evaluated in a larger" . _:b437201153 "2"^^ . _:b437200342 . _:b437201802 . _:b437201152 "2"^^ . _:b437200337 . _:b437201813 . . _:b437201155 "2"^^ . _:b437200336 . _:b437201234 . _:b437201812 . _:b437201154 "2"^^ . _:b437200339 . _:b437201815 . _:b437201165 "2"^^ . _:b437200338 . _:b437201814 . _:b437201164 "2"^^ . _:b437200214 . _:b437200349 . . _:b437201809 . _:b437201167 "2"^^ . . _:b437200348 . . _:b437201808 . . _:b437200604 . _:b437201166 "2"^^ . _:b437200816 . _:b437200351 . _:b437201811 . _:b437201161 "2"^^ . _:b437200289 . _:b437200350 . _:b437201810 . _:b437201160 "2"^^ . . _:b437200345 . _:b437201821 . . . _:b437201163 "2"^^ . _:b437200344 . _:b437201820 . . _:b437201162 "2"^^ . _:b437200347 . _:b437201823 . . _:b437201173 "2"^^ . _:b437200346 . _:b437201603 . _:b437201822 . _:b437201172 "2"^^ . _:b437200357 . _:b437201817 . _:b437200456 . _:b437201175 "2"^^ . _:b437200356 . _:b437201816 . _:b437201670 . _:b437201174 "2"^^ . _:b437200359 . . _:b437201819 . . _:b437200866 . _:b8124659 "Since DSB formation requires only transient expression of ZFNs, gene delivery methods that are suitable to express ZFNs include plasmid DNA nucleofection, integrase-defective lentiviral vectors and adenoviral vectors (>>16<<,19,21). Although non-viral methods such as nucleofection are attractive, this procedure has previously been associated with relatively high rates of toxicity in human CD34+ HSC and loss of engraftment potential (22), although more" . _:b437201169 "2"^^ . . _:b437200358 . . _:b437201818 . _:b437200528 . . _:b437201168 "2"^^ . . _:b437200353 . _:b437201829 . _:b437201171 "2"^^ . _:b437200352 . . _:b437201828 . _:b437201170 "2"^^ . _:b437200355 . _:b437201831 . _:b437201667 . _:b437201181 "2"^^ . _:b437200354 . _:b437200955 . _:b437201830 . _:b437201180 "2"^^ . _:b437200365 . _:b437201220 . _:b437201825 . _:b437201221 . . _:b437201222 . _:b437201183 "2"^^ . _:b437201223 . _:b437200364 . . . _:b437201216 . _:b437201824 . _:b437201217 . _:b437201218 . _:b437201182 "2"^^ . _:b437201219 . _:b437201892 . _:b437200367 . _:b437201228 . _:b437201827 . . _:b437201229 . _:b437200432 . _:b437201177 "2"^^ . _:b437201230 . _:b437201231 . _:b437200366 . _:b437201224 . _:b437201826 . _:b437201256 . . _:b437201225 . _:b437201226 . _:b437201176 "2"^^ . _:b437201227 . _:b437201531 . _:b437200361 . _:b437201236 . _:b437201837 . _:b437201237 . _:b437201238 . _:b437201179 "2"^^ . _:b437200454 . _:b437200360 . _:b437201239 . _:b437201232 . _:b437201836 . _:b437201233 . _:b437201234 . _:b437201178 "2"^^ . _:b437201235 . _:b437200363 . _:b437201244 . _:b437201839 . _:b437200472 . _:b437201245 . _:b437201246 . _:b437201189 "2"^^ . _:b437201087 . . . _:b437201247 . _:b437200362 . _:b437201240 . _:b437201838 . _:b437201241 . _:b437201584 . _:b437201242 . _:b437201188 "2"^^ . _:b437201243 . _:b437200373 . _:b437201252 . _:b437201833 . _:b437201253 . _:b437201254 . _:b437201191 "2"^^ . _:b437201255 . _:b437200372 . _:b437201248 . _:b437201832 . _:b437201249 . _:b437201250 . _:b437201190 "2"^^ . _:b437201251 . _:b437200375 . . _:b437201260 . _:b437201835 . . . _:b437201261 . _:b437201262 . _:b437201185 "2"^^ . _:b437201263 . _:b437200374 . _:b437201256 . _:b437201955 . _:b437201834 . _:b437201257 . _:b437201258 . _:b437201184 "2"^^ . _:b437201259 . _:b437201772 . _:b437200369 . _:b437201268 . _:b437201845 . _:b437201269 . _:b437201270 . _:b437201187 "2"^^ . _:b437201271 . _:b437200368 . _:b437201264 . _:b437201844 . _:b437201265 . . _:b437201266 . _:b437201186 "2"^^ . _:b437201267 . _:b437200371 . _:b437201276 . _:b437201847 . _:b437201277 . . _:b437201197 "2"^^ . _:b437201278 . _:b437201279 . _:b437200370 . _:b437201272 . _:b437201846 . _:b437201273 . _:b437201274 . _:b437201196 "2"^^ . _:b437201275 . _:b437200381 . _:b437201156 . _:b437201841 . _:b437201157 . . _:b437201158 . _:b437201731 . _:b437201199 "2"^^ . . _:b437201159 . _:b437200380 . _:b437201152 . _:b437201840 . _:b437201153 . _:b437201154 . _:b437201198 "2"^^ . . _:b437200852 . . _:b437201155 . _:b437200383 . _:b437201164 . _:b437201843 . _:b437201211 . _:b437201165 . _:b437201166 . _:b437201193 "2"^^ . _:b437201167 . _:b437200382 . _:b437201160 . _:b437201842 . _:b437201161 . _:b437201162 . _:b437201192 "2"^^ . _:b437201163 . _:b437200377 . _:b437201172 . _:b437201853 . _:b437200519 . _:b437201173 . _:b437201174 . _:b437201195 "2"^^ . _:b437201175 . _:b437200376 . _:b437201168 . _:b437201852 . _:b437201169 . _:b437201170 . _:b437201194 "2"^^ . _:b437201171 . _:b437200379 . _:b437201180 . _:b437201855 . _:b437201181 . _:b437201182 . _:b437201205 "2"^^ . _:b437201183 . _:b437200378 . _:b437201176 . _:b437201854 . _:b437201757 . _:b437201177 . _:b437201178 . _:b437201829 . _:b437201204 "2"^^ . _:b437201179 . _:b437201188 . _:b437201849 . . _:b437201189 . _:b437201190 . _:b437201207 "2"^^ . _:b437201191 . _:b437201184 . _:b437201848 . . _:b437201185 . _:b437201186 . _:b437201206 "2"^^ . _:b437201187 . _:b437201196 . _:b437201851 . . _:b437201197 . _:b437201198 . _:b437201201 "2"^^ . _:b437201199 . _:b437201769 . _:b437201457 . _:b437201192 . _:b437201850 . _:b437201594 . _:b437200409 . _:b437201193 . . _:b437201194 . _:b437201200 "2"^^ . _:b437200943 . _:b437201195 . _:b437200601 . _:b437201204 . _:b437201861 . _:b437201205 . _:b437201206 . _:b437201203 "2"^^ . _:b437201207 . _:b437201983 . _:b437201200 . _:b437201860 . _:b437201201 . _:b437201202 . _:b437201202 "2"^^ . _:b437201203 . _:b437201212 . _:b437201863 . _:b437201213 . _:b437201214 . _:b8124716 "In addition, the experience of autologous HSC transplantation in gene therapy for patients with genetic diseases such as ADA-SCID (>>59<<\u201360), chronic granulomatous disease (61) and X-linked adrenoleukodystrophy (62), is that non-myeloablative conditioning can be used to facilitate engraftment of gene-modified autologous HSC with minimal associated toxicity." . _:b437201784 . _:b437201213 "2"^^ . _:b437201215 . _:b437201208 . _:b437201862 . _:b437201209 . _:b437200551 . _:b437201210 . _:b437201212 "2"^^ . _:b437201211 . _:b437201348 . _:b437201857 . _:b437201349 . . _:b437201350 . _:b437201215 "2"^^ . . . _:b437201351 . _:b437201344 . _:b437201856 . _:b437201345 . _:b8124654 "are rapidly repaired by cellular repair pathways, most notably the mutagenic non-homologous end-joining (NHEJ) pathway, which leads to efficient disruption of the gene due to the addition or deletion of nucleotides at the break site (17,>>18<<). A significant advantage of this approach is that permanent gene disruption can occur following only transient expression of the ZFNs, so that a heritable and absolute CCR5-negative phenotype can be achieved with no requirement for the" . . . _:b437201346 . _:b437201214 "2"^^ . _:b437201347 . _:b437201356 . _:b437201859 . _:b437201357 . _:b437201358 . _:b437201258 . _:b437201209 "2"^^ . _:b437201359 . _:b437200976 . _:b437201352 . _:b437201858 . _:b437201353 . _:b437201354 . _:b437201208 "2"^^ . _:b437201355 . _:b437201364 . _:b437201869 . _:b437201808 . _:b437201365 . _:b437201366 . _:b437201211 "2"^^ . _:b437201367 . _:b437201360 . _:b437201868 . _:b437200417 . _:b437201361 . _:b437201617 . _:b437201362 . _:b437201210 "2"^^ . _:b437201363 . _:b437201372 . _:b437201871 . . _:b437201373 . _:b437201374 . _:b437201221 "2"^^ . _:b437201375 . _:b437201095 . _:b437201368 . _:b437201870 . _:b437201369 . _:b437201315 . _:b437201370 . _:b437201220 "2"^^ . . _:b437201371 . _:b437201960 . _:b437201380 . _:b437201865 . _:b437201381 . _:b437201382 . _:b437201223 "2"^^ . _:b437201383 . _:b437201376 . _:b437201864 . _:b437201377 . _:b437200492 . _:b437201378 . _:b437201222 "2"^^ . _:b437201379 . _:b437201388 . _:b437201867 . _:b437201389 . _:b437201390 . _:b437201217 "2"^^ . _:b437201391 . _:b437201384 . _:b437201866 . _:b437201385 . _:b437201386 . _:b437201216 "2"^^ . _:b437201387 . _:b437201396 . _:b437201877 . . _:b437201397 . _:b437201398 . _:b437201219 "2"^^ . _:b437201399 . _:b437201392 . _:b8124642 . _:b437201876 . _:b437201393 . _:b437201394 . _:b437201218 "2"^^ . _:b437201395 . _:b437200997 . _:b437201404 . _:b437201879 . _:b437200265 . _:b437201405 . _:b437201406 . _:b437201229 "2"^^ . _:b437201407 . _:b437201400 . _:b437201878 . _:b437201415 . _:b437201401 . _:b437201402 . _:b437201579 . _:b437201228 "2"^^ . _:b437201403 . . _:b437201284 . _:b437201873 . _:b437201130 . _:b437201285 . _:b437201286 . _:b437201231 "2"^^ . _:b437201287 . _:b437201280 . _:b437201872 . _:b437201281 . _:b437201282 . _:b437201230 "2"^^ . _:b437201283 . _:b437201292 . _:b437201943 . _:b437201875 . _:b437201293 . _:b437201294 . _:b437201836 . _:b437201225 "2"^^ . _:b437201295 . _:b437201288 . _:b437201874 . _:b437201289 . _:b437201290 . _:b437201224 "2"^^ . _:b437201291 . _:b437201300 . _:b437201885 . _:b437201301 . _:b437201302 . _:b437201227 "2"^^ . _:b437201303 . _:b437201296 . _:b437201884 . _:b437201297 . . _:b437201298 . _:b437201226 "2"^^ . . _:b437201299 . _:b437201308 . _:b437201887 . _:b437201309 . _:b437201310 . _:b437201568 . _:b437201237 "2"^^ . . _:b437201311 . _:b437201304 . _:b437201886 . _:b437201305 . _:b437201876 . _:b437201306 . _:b437201236 "2"^^ . _:b437201307 . _:b437201316 . _:b437201881 . _:b437201317 . _:b437201318 . _:b437201239 "2"^^ . _:b437201319 . _:b437201312 . _:b437201880 . _:b437201313 . _:b437201314 . _:b437201238 "2"^^ . . _:b437201315 . . _:b437201324 . _:b437201883 . _:b437201325 . _:b437201326 . _:b437201233 "2"^^ . _:b437201327 . _:b437201320 . _:b437201882 . _:b437201321 . _:b437201736 . _:b437201322 . _:b437201232 "2"^^ . _:b437201323 . _:b437201332 . _:b437201893 . . . _:b437201333 . _:b437201334 . _:b437201235 "2"^^ . _:b437201335 . _:b437201328 . _:b437201892 . _:b437201329 . _:b437201239 . _:b437201330 . _:b437201335 . _:b437201234 "2"^^ . _:b437201331 . _:b437201340 . _:b437201895 . _:b437201341 . . _:b437200558 . _:b437201342 . _:b437201245 "2"^^ . _:b437201343 . _:b437201336 . _:b437201894 . . _:b437201337 . . _:b437201338 . _:b437201244 "2"^^ . _:b437201099 . _:b437201339 . _:b437200964 . _:b437201889 . _:b437200965 . _:b437200966 . _:b437201247 "2"^^ . _:b437200967 . _:b437200960 . _:b437201888 . . _:b437200961 . _:b437200962 . _:b437201246 "2"^^ . _:b437200992 . _:b437200963 . . . _:b437200972 . _:b437201891 . . _:b437200973 . _:b437200974 . _:b437201241 "2"^^ . _:b437200975 . . _:b437200968 . _:b437201890 . _:b437200969 . _:b437200970 . _:b437201240 "2"^^ . _:b437200971 . _:b437200994 . . _:b437200391 . _:b437200980 . _:b437201901 . _:b437200561 . _:b437200981 . _:b437200982 . _:b437201243 "2"^^ . _:b437200983 . _:b437201516 . _:b437200976 . _:b437201900 . _:b437201257 . _:b437201240 . _:b437200977 . _:b437201735 . _:b437200978 . _:b437201242 "2"^^ . _:b437200979 . _:b437200988 . _:b437201903 . _:b437200989 . . _:b437201253 "2"^^ . _:b437200990 . _:b437200991 . _:b437200881 . _:b437200984 . _:b437201902 . _:b437200985 . _:b437201475 . . _:b437200986 . _:b437201252 "2"^^ . _:b437200987 . _:b437200400 . _:b437200996 . _:b8124664 "Cel 1 analysis was used to assess the levels of CCR5 disruption since only a minority of human CD34+ HSC express CCR5 (>>26<<), so that it is not possible to monitor the effects of this modification by FACS analysis of the population." . _:b437201897 . _:b437200997 . _:b437200998 . _:b437201255 "2"^^ . _:b437200999 . _:b437201179 . _:b437200992 . _:b437201896 . . _:b437200993 . _:b437200357 . _:b437200275 . _:b437200994 . _:b437201254 "2"^^ . _:b437200995 . _:b437201004 . _:b437201899 . _:b437201005 . _:b437201006 . _:b437201249 "2"^^ . _:b437200445 . . _:b437201007 . _:b437201000 . _:b437201898 . . _:b437201001 . . _:b437201002 . _:b437201248 "2"^^ . _:b437201003 . _:b437201012 . _:b437201909 . _:b437201013 . _:b437201014 . _:b437201251 "2"^^ . _:b437201015 . _:b437201008 . _:b437201908 . _:b437201419 . _:b437201314 . _:b437201009 . _:b437201010 . _:b437201728 . _:b437201428 . _:b437201250 "2"^^ . _:b437201011 . _:b437201872 . _:b437201454 . _:b437201080 . _:b437201020 . _:b437201911 . _:b437201021 . _:b437201022 . _:b437201261 "2"^^ . _:b437201023 . _:b437201016 . _:b437201910 . _:b437201017 . _:b437201018 . _:b437201260 "2"^^ . . . _:b437201019 . _:b437201195 . _:b437200900 . _:b437201905 . _:b437200901 . _:b437200902 . _:b437201263 "2"^^ . . _:b437200903 . . _:b437200896 . _:b437201904 . _:b437200897 . _:b437200898 . _:b437201262 "2"^^ . _:b437200899 . _:b437200908 . _:b437201907 . _:b437200909 . _:b437200910 . _:b437201257 "2"^^ . _:b437200911 . _:b437200904 . _:b437201906 . _:b437200905 . _:b437201449 . _:b437200906 . _:b437201256 "2"^^ . _:b437200907 . . _:b437200916 . _:b437201917 . _:b437200917 . . . _:b437200918 . _:b437201259 "2"^^ . _:b437200919 . _:b437200912 . _:b437201916 . _:b437200913 . _:b437200914 . _:b437201928 . _:b437201258 "2"^^ . _:b437200915 . _:b437200924 . _:b437201919 . _:b437200925 . . _:b437200926 . _:b437201269 "2"^^ . _:b437201079 . . _:b437200927 . _:b437200920 . _:b437201918 . _:b437200921 . _:b437200922 . _:b437201268 "2"^^ . . _:b437200923 . _:b437200932 . _:b437201913 . _:b437200933 . _:b437200934 . _:b437201271 "2"^^ . _:b437200935 . _:b437200928 . _:b437201912 . _:b437200929 . _:b437200930 . _:b437201270 "2"^^ . _:b437200707 . _:b437200931 . _:b437200940 . _:b437201915 . _:b437200656 . _:b437200785 . _:b437200941 . _:b437200942 . _:b437201265 "2"^^ . _:b437200943 . _:b437200936 . _:b437201914 . _:b437200937 . _:b437200938 . _:b437201264 "2"^^ . _:b437200939 . _:b437200948 . _:b437201413 . _:b437200949 . _:b437200950 . _:b437201267 "2"^^ . _:b437200951 . . _:b437200944 . _:b437201412 . _:b437200945 . _:b437200946 . _:b437201699 . _:b437201266 "2"^^ . _:b437200947 . _:b437200956 . _:b437201415 . _:b437200957 . _:b437200958 . _:b437201277 "2"^^ . _:b437200959 . _:b437200952 . _:b437201414 . _:b437200953 . _:b437200748 . _:b437200954 . _:b437201276 "2"^^ . _:b437200955 . _:b437201092 . _:b437201409 . _:b437201093 . _:b437201094 . _:b437201279 "2"^^ . . _:b437201095 . _:b437201088 . _:b437201408 . _:b437201089 . _:b437201090 . _:b437201278 "2"^^ . _:b437201091 . _:b437201100 . _:b437201411 . _:b437201101 . _:b437201102 . _:b437201273 "2"^^ . _:b437201103 . _:b437201096 . _:b437201410 . _:b437201097 . _:b437200249 . . . _:b437201098 . _:b437201272 "2"^^ . _:b437201099 . _:b437201108 . _:b437201421 . _:b437201109 . _:b437201110 . _:b437201275 "2"^^ . _:b437201111 . _:b437201104 . _:b437201420 . _:b437201105 . . _:b437201106 . _:b437201274 "2"^^ . _:b437201107 . _:b437200576 . _:b437201116 . _:b437201423 . _:b437201117 . _:b437201118 . _:b437201285 "2"^^ . _:b437201119 . _:b437201112 . _:b437201422 . _:b437201113 . _:b437201114 . _:b437201284 "2"^^ . _:b437201115 . _:b437201124 . _:b437201727 . _:b437201417 . _:b437201125 . _:b437201126 . _:b437201287 "2"^^ . _:b437201127 . _:b437201110 . _:b437201054 . _:b437201120 . _:b437201416 . _:b437201121 . . _:b437200556 . _:b437201122 . _:b437201286 "2"^^ . _:b437201123 . _:b437201132 . _:b437201419 . _:b437200878 . _:b437201133 . . _:b437201134 . _:b437201281 "2"^^ . _:b437201135 . _:b437201128 . _:b437201418 . _:b437201129 . _:b437201130 . _:b437201280 "2"^^ . _:b437201238 . _:b437201131 . _:b437201140 . _:b437201429 . _:b437201141 . _:b437201142 . _:b437201283 "2"^^ . _:b437201143 . _:b437201136 . _:b437201428 . _:b437201137 . _:b437201138 . _:b437201282 "2"^^ . _:b437201139 . _:b437201148 . _:b437201431 . _:b437201149 . _:b437201150 . _:b437201293 "2"^^ . _:b437201007 . _:b437201151 . . _:b437201144 . _:b437201430 . _:b437201145 . _:b437201146 . _:b437201292 "2"^^ . _:b437201147 . _:b437201427 . _:b437201028 . _:b437201425 . _:b437201280 . _:b437201029 . _:b437200683 . _:b437201030 . _:b437201295 "2"^^ . _:b437201031 . _:b437201024 . _:b437201424 . _:b437200966 . _:b437201025 . _:b437201026 . _:b437201294 "2"^^ . _:b437201027 . . . _:b437201036 . _:b437201427 . _:b437201037 . _:b437201038 . _:b437201289 "2"^^ . _:b437201039 . _:b437201032 . _:b437201426 . _:b437201033 . . _:b437201034 . _:b437201288 "2"^^ . _:b437201035 . _:b437201044 . _:b437201437 . _:b437201045 . _:b437201046 . _:b8124645 . _:b437201413 . _:b437201291 "2"^^ . . _:b437201047 . . _:b437201040 . _:b437201436 . _:b437201041 . _:b437201042 . _:b437201290 "2"^^ . _:b437201043 . _:b437201052 . _:b437201439 . _:b437201053 . _:b437201300 . _:b437201054 . _:b437201301 "2"^^ . . _:b437201055 . _:b437201048 . _:b437201438 . _:b437201049 . _:b437200970 . _:b437201050 . _:b437201300 "2"^^ . _:b437201051 . _:b437201060 . _:b437201433 . _:b437201061 . _:b437201062 . _:b437201303 "2"^^ . _:b437201063 . _:b437201998 . _:b437201056 . _:b437201432 . _:b437201057 . _:b437200237 . _:b437201058 . _:b437201302 "2"^^ . _:b437201059 . _:b8124727 "in certain sanctuary sites, even in well-controlled patients on ART (43,68), and this could provide a low level of selection, although drug intensification trials have not provided evidence of any such ongoing HIV-1 replication (>>69<<). It is also possible that the high levels of CCR5 disruption we achieved without selection, if extrapolated to patients, could be sufficient to provide a therapeutic effect even in the absence of a strong selective pressure." . _:b437201517 . _:b437200494 . _:b437201919 . _:b437201435 . _:b437201068 . _:b437201069 . _:b437201070 . _:b437201297 "2"^^ . . _:b437201071 . _:b8124721 "in patients treated with CCR5 inhibitors unless pre-existing CXCR4-tropic viruses were present before therapy, and resistance to the drugs can occur, instead, because of viral adaptations to the drug-bound form of the CCR5 co-receptor (>>66<<,67). Interestingly, the patient receiving the CCR5\u039432 HSC transplantation harbored CXCR4-utilizing viruses before the procedure, yet he did not experience a tropism switch, and instead, successfully controlled his HIV-1 infection" . _:b437201064 . _:b437201434 . _:b437201065 . _:b437201066 . _:b437201296 "2"^^ . _:b437201106 . _:b437201067 . _:b437201076 . _:b437201445 . _:b437201077 . . _:b437201078 . _:b437201299 "2"^^ . _:b437201079 . _:b437201072 . _:b437201444 . _:b437201073 . _:b437201074 . _:b437201298 "2"^^ . _:b437201075 . _:b437201664 . _:b437201084 . _:b8124675 "These observations mimic the human infection, where depletion of CD4+ CCR5+ lymphocytes results from a combination of direct infection, systemic immune activation and the upregulation of CCR5 on thymocytes (36,>>37<<,38). Following infection, blood samples were collected from the mice every two weeks and analyzed for HIV-1 RNA levels, T cell subsets and the extent of CCR5 disruption. At 8\u201312 weeks post-infection, animals were euthanized and necropsy" . _:b437201447 . _:b437201085 . _:b437201086 . _:b437201309 "2"^^ . _:b437201087 . _:b437201080 . _:b437201446 . _:b437201081 . . . _:b437201082 . _:b437201308 "2"^^ . _:b437201083 . . _:b437201732 . _:b437201441 . _:b437200512 . _:b437201733 . _:b437201734 . _:b437201311 "2"^^ . _:b437201735 . _:b437201728 . _:b437201440 . _:b437201729 . _:b437201730 . _:b437201310 "2"^^ . . _:b437201731 . _:b437201740 . _:b437201443 . _:b437201741 . _:b437201742 . _:b437201791 . _:b437201305 "2"^^ . _:b437201743 . _:b437201736 . _:b437201442 . _:b437201737 . . _:b437201738 . _:b437201304 "2"^^ . _:b437201739 . _:b8124711 "feature of HIV-1 replication in mucosal tissues is an ongoing cycle of T cell death and the accompanying recruitment of replacement T cells, whose activated state means that they are also highly permissive targets for HIV-1 infection (>>37<<). This is especially true in the GALT, which represents a key battleground in HIV-1 infection (54\u201356)." . _:b437201748 . _:b437201453 . _:b437201749 . . _:b437201750 . _:b437201307 "2"^^ . _:b437201056 . _:b437201751 . _:b437200250 . _:b437201744 . _:b437201452 . _:b437201745 . _:b437201746 . _:b437201641 . _:b437201306 "2"^^ . _:b437201747 . _:b437201756 . _:b437201455 . _:b437201757 . _:b437201758 . _:b437201317 "2"^^ . . _:b437201759 . _:b437201752 . _:b437201454 . _:b437201753 . _:b437201754 . _:b437201316 "2"^^ . . . . _:b437201755 . _:b437201764 . _:b437201449 . . _:b437200402 . _:b437201765 . _:b437201766 . _:b437201319 "2"^^ . _:b437201767 . _:b437201760 . _:b437201448 . _:b437201761 . _:b437201762 . _:b437201318 "2"^^ . _:b437201763 . _:b437201772 . _:b437201451 . _:b437201773 . _:b437201774 . _:b437201313 "2"^^ . _:b437201775 . _:b437201768 . _:b437201450 . _:b437201769 . . _:b437201312 "2"^^ . _:b437201770 . _:b437201771 . _:b437201780 . _:b437201662 . _:b437201461 . _:b437201781 . _:b437201782 . _:b437201315 "2"^^ . _:b437201128 . _:b437201783 . _:b437201776 . _:b437201460 . _:b437201777 . . _:b437201778 . _:b437201314 "2"^^ . _:b437201226 . . _:b437201779 . _:b437201788 . _:b437201463 . _:b437201789 . _:b437201790 . _:b437201325 "2"^^ . _:b437201791 . _:b437201366 . . _:b437201784 . _:b437201462 . _:b437201785 . _:b437201786 . _:b437201324 "2"^^ . _:b437201787 . _:b437201668 . _:b437201457 . _:b437200236 . _:b437201669 . _:b437201670 . _:b437201327 "2"^^ . _:b437201671 . _:b437201664 . _:b437201456 . _:b437200858 . _:b437201665 . _:b437201666 . _:b437201326 "2"^^ . _:b437201667 . _:b437201794 . _:b437201125 . _:b437200777 . _:b437201676 . _:b437201459 . _:b437201677 . . _:b437201678 . _:b437201321 "2"^^ . _:b437201679 . _:b437200412 . _:b437201672 . _:b437201458 . _:b437201673 . _:b437201674 . _:b437201320 "2"^^ . _:b437201675 . _:b437201684 . _:b437201469 . . _:b437201685 . _:b437201686 . _:b437201352 . _:b437201323 "2"^^ . _:b437201687 . _:b437201680 . _:b437201468 . _:b437201681 . _:b437201325 . _:b437201682 . _:b437201322 "2"^^ . _:b437201683 . _:b437201692 . _:b437201471 . _:b437201693 . . _:b437201694 . _:b437201333 "2"^^ . _:b437201695 . _:b437201196 . _:b437201688 . _:b437201470 . _:b437201689 . _:b437200848 . _:b437201690 . _:b437201332 "2"^^ . _:b437201691 . _:b437201700 . _:b437201465 . _:b437201701 . _:b437201702 . _:b437201335 "2"^^ . _:b437201703 . _:b437201115 . _:b437201696 . _:b437201464 . _:b437201697 . _:b437201698 . _:b437201334 "2"^^ . _:b437201699 . _:b437201708 . _:b437201467 . _:b437201709 . _:b437201710 . _:b437201329 "2"^^ . _:b437201711 . . . _:b437201466 . _:b437201704 . _:b437201705 . _:b437201706 . _:b437201328 "2"^^ . _:b437201707 . _:b437201716 . _:b437201477 . _:b437201717 . _:b437201483 . _:b437201718 . _:b437201331 "2"^^ . _:b437201719 . _:b437201712 . _:b437201476 . _:b437201713 . _:b437201714 . _:b437201330 "2"^^ . _:b437201715 . _:b437201724 . _:b437201479 . . _:b437201725 . _:b437201726 . _:b437201341 "2"^^ . _:b437201727 . _:b437200567 . . _:b437201720 . _:b437201478 . _:b437201721 . _:b437201722 . _:b437201340 "2"^^ . _:b437201723 . _:b437201860 . _:b437201473 . _:b437201861 . . . _:b437201343 "2"^^ . _:b437201862 . . _:b437201863 . _:b437201856 . _:b437201472 . . _:b437200277 . _:b437201857 . . _:b437201858 . _:b437201342 "2"^^ . _:b437201859 . _:b437201868 . _:b437201475 . _:b437201869 . _:b437201870 . _:b437201337 "2"^^ . _:b437201871 . _:b437201864 . _:b437201474 . _:b437201865 . _:b437201866 . _:b437201336 "2"^^ . . _:b437201867 . _:b437201876 . _:b437201485 . _:b8124696 "discussion" . _:b437201877 . _:b437201878 . _:b437201339 "2"^^ . . _:b437201879 . _:b437201872 . _:b437201484 . _:b437201873 . _:b437201874 . _:b8124720 "CXCR4-tropic viruses have been associated with accelerated disease progression (>>65<<), so that selection for such strains could be an undesirable consequence of targeting CCR5." . _:b437201338 "2"^^ . _:b437201875 . . _:b437201487 . _:b437201884 . _:b437201885 . _:b8124650 "The targeted cell populations include both mature peripheral T cells or HSC, and the loss of CCR5 in HSC appears to have no adverse effects on hematopoiesis (12,>>13<<,15)." . _:b437201886 . _:b437201349 "2"^^ . . . _:b437200308 . _:b437201887 . _:b437201880 . _:b437201486 . _:b437200851 . . _:b437201881 . _:b437201882 . _:b437201348 "2"^^ . _:b437201883 . _:b437201892 . _:b437201481 . _:b437201893 . _:b437201973 . _:b437201894 . _:b437201351 "2"^^ . _:b437201895 . _:b437201888 . _:b437201480 . . . _:b437201889 . _:b437201890 . _:b437201350 "2"^^ . _:b437201891 . . . _:b437201900 . _:b437201483 . _:b437201901 . _:b437201902 . _:b437201345 "2"^^ . _:b437201903 . _:b437201036 . _:b437201896 . _:b437201482 . _:b437201897 . _:b437200217 . _:b437201344 "2"^^ . _:b437201898 . . _:b437201899 . _:b437201421 . _:b437201908 . _:b437201493 . _:b437201909 . _:b437200810 . _:b437201910 . _:b437201347 "2"^^ . _:b437201911 . _:b437201904 . _:b437201492 . . _:b437201905 . . _:b437201346 "2"^^ . _:b437201906 . _:b437201907 . . _:b437201495 . _:b437201916 . _:b437201917 . _:b437201918 . _:b437201357 "2"^^ . _:b437201919 . _:b437201912 . _:b437201494 . _:b437201913 . . _:b437201914 . _:b8124692 "Tissue samples were manually agitated in PBS before filtering through a sterile 70 \u03BCm nylon mesh screen (Fisher Scientific) and suspension cell preparations produced as previously described (>>19<<). Intestinal samples were processed as previously described (73), with the modification that the mononuclear cell population was isolated following incubation in citrate buffer and collagenase enzyme for 2 hrs, followed by nylon wool" . _:b437201356 "2"^^ . . _:b437201915 . _:b437201796 . _:b437201489 . _:b437200886 . _:b437201797 . _:b437201798 . _:b437201359 "2"^^ . _:b437200868 . _:b437201799 . _:b437200660 . _:b437200652 . _:b437201488 . . . _:b437201792 . _:b437201793 . _:b437201794 . _:b437201358 "2"^^ . _:b437201795 . _:b437201804 . _:b437201491 . _:b437200828 . _:b437201805 . _:b437201806 . _:b437201353 "2"^^ . _:b437201807 . _:b437201800 . _:b437201490 . _:b437201801 . _:b437201802 . _:b437201352 "2"^^ . _:b437201803 . _:b437201812 . _:b437201501 . _:b437201813 . _:b437201814 . _:b437201355 "2"^^ . _:b437201815 . . _:b437201808 . _:b437201500 . . _:b437201809 . _:b437200483 . _:b437201810 . _:b437201354 "2"^^ . _:b437201811 . . _:b437201820 . _:b437201503 . _:b437201821 . _:b437201822 . _:b437201365 "2"^^ . _:b437201823 . _:b437201816 . _:b437201502 . _:b437201817 . _:b437201818 . _:b437201364 "2"^^ . _:b437201819 . _:b437200817 . _:b437201828 . _:b437201497 . _:b437201829 . _:b437201830 . _:b437201367 "2"^^ . _:b437201831 . _:b437201824 . _:b437201496 . . _:b437200315 . _:b437201825 . _:b437201826 . _:b437201366 "2"^^ . . _:b437201827 . . _:b437201836 . _:b437201499 . _:b437201837 . _:b437201838 . _:b437201361 "2"^^ . _:b437201839 . _:b437201832 . _:b437201498 . . _:b437201833 . _:b437201834 . _:b437201957 . _:b437201360 "2"^^ . _:b437201835 . _:b437201844 . _:b437201509 . _:b437201845 . . _:b437201363 "2"^^ . _:b437201846 . _:b437201847 . _:b437201840 . _:b437201508 . _:b437201841 . _:b437201016 . _:b437201842 . _:b437201362 "2"^^ . _:b437201843 . _:b437201852 . _:b437201511 . _:b437201853 . _:b437201854 . _:b437201373 "2"^^ . _:b437201855 . . _:b437201848 . _:b437201510 . _:b437200538 . _:b437201849 . . . _:b437201850 . _:b437201372 "2"^^ . . _:b437201851 . _:b437201476 . _:b437201505 . _:b437201477 . _:b437201478 . _:b437201375 "2"^^ . _:b437201479 . _:b437201472 . _:b437201827 . _:b437201504 . _:b437201473 . _:b437201474 . _:b437201374 "2"^^ . _:b437201475 . _:b437201484 . _:b437201507 . _:b437201485 . _:b437201486 . _:b437201369 "2"^^ . _:b437201487 . . _:b437201480 . _:b437201506 . _:b437201481 . _:b437201482 . _:b437201368 "2"^^ . _:b437201483 . . _:b437201492 . _:b437201517 . . . _:b437201493 . _:b437201494 . _:b437201371 "2"^^ . . . _:b437201495 . . . _:b437201488 . _:b437201516 . _:b437201489 . _:b437201490 . _:b437201370 "2"^^ . . . _:b437201802 . _:b437201491 . _:b437201500 . _:b437201519 . _:b437200258 . _:b437201501 . _:b437201502 . _:b437201381 "2"^^ . _:b437200368 . _:b437201503 . _:b437201496 . _:b437201518 . _:b437201497 . _:b437201197 . _:b437201498 . _:b437201380 "2"^^ . _:b437201499 . _:b437201508 . _:b437201513 . _:b437201236 . _:b437201509 . _:b437201510 . _:b437201383 "2"^^ . _:b437201511 . _:b437201504 . _:b437201512 . _:b437201505 . _:b437201506 . _:b437201382 "2"^^ . _:b437201507 . _:b437200911 . _:b437201516 . _:b437201515 . _:b437201517 . _:b437201518 . _:b437201377 "2"^^ . _:b437201519 . _:b437201512 . _:b437201514 . _:b437201513 . _:b437201514 . _:b437201376 "2"^^ . _:b437201515 . . _:b437201524 . _:b437201525 . _:b437201525 . _:b437201526 . _:b437201379 "2"^^ . _:b437201527 . _:b437201520 . _:b437201524 . _:b437201521 . _:b437201522 . _:b437201708 . _:b437201378 "2"^^ . _:b437200532 . _:b437201523 . _:b437201532 . _:b437201527 . _:b437201533 . _:b437201534 . _:b437201389 "2"^^ . _:b437201535 . _:b437201528 . _:b437201526 . _:b437201529 . _:b437201530 . _:b437201388 "2"^^ . _:b437201531 . _:b437201412 . _:b437201521 . _:b437201413 . _:b437201414 . _:b437201391 "2"^^ . _:b437201415 . _:b437201408 . _:b437201520 . _:b437201409 . _:b437201410 . _:b437201390 "2"^^ . _:b437201411 . _:b437201420 . _:b437201523 . _:b437201421 . _:b437201422 . _:b437201385 "2"^^ . _:b437201423 . _:b437201416 . _:b437201522 . _:b437201417 . _:b437201418 . _:b437201384 "2"^^ . _:b437201419 . _:b437201121 . _:b437201428 . _:b437201533 . _:b437201429 . _:b437201430 . _:b437201387 "2"^^ . _:b437201431 . _:b437201424 . _:b437201532 . _:b437201425 . _:b437201426 . _:b437201386 "2"^^ . _:b437201427 . _:b437201436 . _:b437201535 . _:b437201437 . _:b437201438 . _:b437201397 "2"^^ . _:b437201439 . _:b437201432 . _:b437201534 . _:b437201433 . _:b437201434 . _:b437201396 "2"^^ . _:b437201435 . _:b437201444 . _:b437201529 . . _:b437201445 . _:b437201643 . _:b437201446 . _:b437201399 "2"^^ . _:b437201447 . _:b437201440 . _:b437201528 . _:b437201441 . _:b437201442 . _:b437201398 "2"^^ . _:b437201443 . . _:b437201452 . _:b437201531 . _:b437201453 . _:b437201454 . _:b437201393 "2"^^ . _:b437201455 . _:b437200741 . _:b437201448 . _:b437201530 . _:b437201449 . _:b437201450 . _:b437201392 "2"^^ . _:b437201451 . _:b437200301 . _:b437201460 . _:b437201541 . _:b437201461 . _:b437201462 . _:b437201395 "2"^^ . _:b437200460 . _:b437201463 . _:b437201456 . _:b437201540 . _:b437201457 . _:b437201458 . _:b437201394 "2"^^ . _:b437200797 . _:b437201459 . _:b437201468 . _:b437201543 . _:b437201469 . . _:b437201470 . _:b437201405 "2"^^ . _:b437201471 . _:b437201464 . _:b437201542 . _:b437201465 . _:b437201466 . _:b437201404 "2"^^ . _:b437201467 . _:b437201604 . _:b437201537 . _:b437201605 . _:b437201606 . _:b437201407 "2"^^ . _:b437201607 . _:b437201600 . _:b437201536 . . . _:b437201601 . . _:b437200738 . _:b437201406 "2"^^ . _:b437201602 . _:b437201603 . _:b437201612 . _:b437201539 . . _:b437201613 . _:b437201614 . _:b437201401 "2"^^ . . _:b437201615 . _:b437201608 . _:b437201538 . _:b437201609 . _:b437201610 . _:b437201400 "2"^^ . _:b437201611 . _:b437201813 . _:b437201620 . _:b437201549 . _:b437201621 . _:b437201622 . _:b437201403 "2"^^ . _:b437201623 . . _:b437201616 . _:b437201548 . . _:b437201617 . _:b437201618 . _:b437201402 "2"^^ . _:b437201619 . _:b437201628 . _:b437201551 . _:b437200850 . _:b437201629 . _:b437201630 . _:b437201631 . _:b437200637 . . _:b437201550 . _:b437201624 . . _:b437201625 . _:b437201626 . _:b437201627 . _:b437201636 . _:b437201796 . _:b437201545 . _:b437201637 . _:b437201638 . _:b437201639 . _:b437201632 . _:b437201544 . _:b437201633 . . . _:b437201634 . _:b437201635 . . _:b437201547 . _:b437201644 . _:b437201645 . _:b437201646 . _:b437201647 . . _:b437201546 . _:b437201640 . _:b437201641 . _:b437201642 . _:b8124653 "are rapidly repaired by cellular repair pathways, most notably the mutagenic non-homologous end-joining (NHEJ) pathway, which leads to efficient disruption of the gene due to the addition or deletion of nucleotides at the break site (>>17<<,18). A significant advantage of this approach is that permanent gene disruption can occur following only transient expression of the ZFNs, so that a heritable and absolute CCR5-negative phenotype can be achieved with no requirement for" . _:b437201473 . _:b437201643 . _:b437201652 . _:b437201557 . _:b437201653 . _:b437200811 . _:b437201654 . _:b437201655 . _:b437201648 . _:b437201556 . _:b437201649 . . _:b437201650 . _:b437201651 . _:b437201660 . _:b437201559 . _:b437201661 . . _:b437201662 . _:b437201663 . _:b437201656 . _:b437201558 . _:b437201219 . _:b437201657 . _:b8124668 "adverse effects on cell viability resulting from the nucleofection procedure may be offset by the high levels of CCR5 disruption achieved, as well as the speed and simplicity of the procedure when compared to viral vector systems (19,>>21<<)." . _:b437201658 . _:b437201659 . _:b437201540 . _:b437201553 . _:b437200466 . _:b437201541 . _:b437201542 . _:b437201482 . _:b437201543 . _:b437201536 . _:b437201552 . _:b437200509 . _:b437201537 . _:b437201538 . _:b437201539 . . _:b437201548 . _:b437201555 . _:b437201549 . _:b437201550 . _:b437201551 . . _:b437201544 . _:b437201554 . _:b437201545 . _:b437201546 . _:b437201547 . _:b437201225 . _:b437201556 . _:b437201565 . _:b437201557 . _:b437201558 . _:b437201559 . _:b437201552 . _:b437201564 . _:b437201553 . _:b437201554 . _:b437201555 . _:b437201564 . _:b437201567 . _:b437201565 . . _:b437201566 . _:b437200645 . . _:b437200534 . _:b437201567 . _:b437201560 . _:b437201566 . . _:b437201561 . _:b437201562 . . _:b437201563 . _:b437201572 . _:b437201561 . _:b437201573 . _:b8124684 . _:b437201574 . _:b437201575 . _:b437201687 . _:b437201568 . _:b437201560 . _:b437201444 . _:b437201569 . _:b437201570 . _:b437201571 . _:b437201580 . _:b437201563 . _:b437201581 . _:b437201582 . _:b437201583 . _:b437201576 . _:b437201562 . _:b437201111 . . _:b437201577 . _:b437201578 . _:b437200819 . _:b437201579 . _:b437201588 . _:b8124662 . _:b437201573 . _:b437201589 . _:b437201590 . . _:b437201591 . . _:b437201584 . _:b437201572 . _:b437201585 . _:b437201586 . _:b437201587 . _:b437201596 . _:b437201575 . _:b437201597 . _:b437201598 . _:b437201599 . _:b437201592 . _:b437201574 . _:b437201593 . _:b437201594 . _:b437201595 . _:b437201569 . _:b437201552 . . _:b437201568 . . _:b437201571 . . _:b437201570 . _:b437201912 . _:b8124703 . _:b437201581 . _:b437201580 . . _:b8124680 "Depletion of both single and double positive thymocytes has been reported to occur as a consequence of the up-regulation of CCR5 on these cells following HIV-1 infection (36,>>38<<), and likely contributed to both the observed depletion of the thymus, as well as a reduction in the numbers of mature CD4+ and CD8+ T cells observed in other tissues." . _:b437201463 . . _:b437201583 . . . . _:b437201582 . . _:b437201729 . . _:b437201577 . . _:b437201576 . _:b8124646 . _:b437201579 . _:b437200461 . _:b437201578 . _:b437201255 . _:b437201589 . . . _:b437201588 . . . _:b437200557 . _:b437201591 . _:b437200399 . . . _:b437201590 . _:b437201456 . _:b437201138 . _:b437201585 . _:b437201480 . _:b437201584 . _:b437201587 . _:b8124724 "the patient receiving the CCR5\u039432 HSC transplantation harbored CXCR4-utilizing viruses before the procedure, yet he did not experience a tropism switch, and instead, successfully controlled his HIV-1 infection long-term (9,>>10<<). For the present, similar to the recommendations for the use of CCR5 inhibitor drugs, it may be prudent to restrict CCR5 ZFN treatment of HSC to individuals with no detectable CXCR4-tropic viruses." . _:b437201586 . . _:b437201962 . _:b437201597 . . _:b437201596 . . . . _:b437201909 . . _:b437201599 . _:b437200600 . . . . _:b437201598 . _:b437201144 . . _:b437201593 . _:b437201592 . _:b8124647 "However, the risks associated with HSC transplantation, as well as the impracticality of obtaining sufficient numbers of matched CCR5\u039432 donors (>>10<<), mean that the more widespread use of this approach will require methods that allow the effective and safe generation of CCR5-negative cells in a patient\u2019s own HSC." . _:b437201174 . _:b437201595 . _:b437200549 . _:b437200186 . _:b437200496 . _:b437201594 . _:b437200342 . . _:b437201270 . _:b437201605 . . . . _:b437201604 . . _:b437201607 . _:b437201606 . _:b437201580 . _:b437201601 . _:b437201073 . _:b8124707 . . _:b437201600 . . _:b437201603 . _:b437200457 . . . _:b437201602 . . . _:b437201613 . _:b437200596 . . . . _:b437201612 . _:b437200247 . _:b437201615 . . _:b437201614 . . _:b437201349 . _:b437201609 . _:b437201467 . _:b437200782 . . _:b437201608 . . _:b437201611 . . . _:b437201610 . _:b437200631 . _:b437201621 . _:b437201864 . _:b437201935 . _:b437201494 . _:b437201503 . _:b437201620 . . _:b437201871 . _:b437201623 . . . _:b437201622 . . . _:b437201617 . _:b437201616 . . _:b437201003 . . . _:b437201619 . . _:b437201071 . _:b437201618 . _:b437200692 . _:b437201629 . . . _:b437201628 . . _:b437201233 . _:b437201316 . _:b437201631 . _:b437201630 . _:b437201625 . _:b437201624 . _:b437201627 . _:b437201626 . _:b437201637 . _:b437201636 . . _:b437201639 . _:b437201638 . _:b437201715 . _:b437201988 . _:b437201633 . _:b437201989 . _:b437201472 . _:b437201990 . . _:b437201991 . _:b437201984 . _:b437201632 . _:b437201985 . _:b437201986 . _:b437201987 . _:b437201996 . _:b437201635 . _:b437201997 . . _:b437201998 . _:b437201999 . _:b437201992 . _:b437201634 . _:b437201993 . _:b437201982 . _:b437201994 . _:b437201995 . . _:b437202004 . _:b437201645 . . _:b437202005 . _:b437202006 . _:b437202007 . _:b437202000 . _:b437201644 . . . _:b437202001 . . _:b437202002 . _:b437202003 . _:b437201647 . . _:b437202008 . _:b437201646 . _:b437201641 . . _:b437200849 . _:b437201640 . . . _:b437201643 . _:b437201269 . . _:b437201642 . . _:b437201653 . _:b437200563 . _:b437201652 . _:b8124706 "Off-target events were only observed at significant levels at the homologous CCR2 locus (>>19<<). Mouse studies have not found any deleterious phenotype associated with loss of CCR2 (48), and human genetic studies have even suggested a beneficial phenotype from the loss of this gene in HIV-infected individuals (49). Although not" . _:b437201655 . _:b437201191 . . . _:b437201654 . . _:b437201924 . _:b437201649 . _:b437201925 . _:b437201926 . _:b437201927 . _:b437201920 . _:b437201648 . _:b437201921 . _:b437201922 . _:b437201923 . _:b437201932 . _:b437201651 . _:b437201933 . _:b437201934 . _:b437201935 . _:b437200830 . _:b437201928 . _:b437201650 . _:b437201120 . _:b437201929 . _:b437201930 . _:b437201931 . _:b437201940 . _:b437201661 . _:b437200888 . _:b437201941 . _:b437201942 . _:b437201943 . _:b437201242 . _:b437200281 . _:b437201936 . _:b437201660 . _:b437201937 . . _:b437201938 . _:b437200375 . _:b437201939 . _:b437200704 . _:b437201663 . . _:b437201948 . _:b437201949 . _:b437201950 . _:b437201951 . _:b437201944 . _:b437201662 . _:b437201945 . _:b437201336 . _:b437200630 . _:b437201946 . _:b437201947 . _:b437201956 . _:b437201657 . . _:b437201957 . . _:b437201958 . _:b437200285 . . _:b437201959 . _:b437201952 . _:b437201656 . _:b437201953 . _:b437201954 . _:b437200845 . _:b437201782 . _:b437201955 . _:b437201964 . _:b437201659 . _:b437201965 . . _:b437201966 . _:b437200990 . _:b437201967 . _:b437201960 . _:b437201658 . . _:b437201961 . _:b437201962 . _:b437201963 . _:b437201544 . _:b437201043 . . _:b437201972 . _:b437201157 . _:b437201973 . _:b437201974 . _:b437201975 . . _:b437201968 . _:b437201156 . _:b437201969 . _:b437201970 . _:b437201971 . _:b437201980 . _:b437201159 . . _:b437201981 . _:b437200423 . . _:b437201982 . _:b437201983 . _:b437200867 . _:b437201976 . _:b437201158 . _:b437200403 . _:b437201977 . _:b437201405 . _:b437201978 . _:b437201979 . _:b437201153 . _:b437201970 . . _:b437201152 . _:b437201155 . _:b437201706 . _:b437201499 . . _:b437201154 . _:b437201165 . _:b437200898 . _:b437201686 . . . _:b437200324 . _:b437201164 . . . _:b437201167 . _:b437200421 . _:b437201549 . _:b437201166 . . . _:b437201161 . _:b437201160 . . . _:b437201163 . _:b437201276 . . _:b437201162 . _:b437201533 . . _:b437201173 . . _:b437200723 . _:b437201172 . _:b437200297 . _:b437200322 . _:b437201175 . . _:b437201174 . . _:b437201169 . _:b437201262 . _:b437201168 . _:b437201513 . . _:b437201171 . _:b437201170 . . _:b437201181 . _:b437201625 . _:b437201180 . _:b437201222 . . _:b437201183 . _:b437200784 . . _:b437201182 . . _:b437201249 . _:b437201177 . . _:b437201176 . _:b437200334 . _:b437201179 . _:b437201178 . _:b437201811 . _:b437201189 . _:b437200622 . . _:b437201188 . _:b437201191 . _:b8124681 "We used both the Cel 1 heteroduplex mismatch assay and a specific PCR amplification that detects a 5bp duplication at the ZFN target site that typically accounts for 10\u201330% of total modifications induced by the CCR5 ZFNs (>>19<<). Both assays revealed rapid selection for ZFN-disrupted alleles, reaching the upper limit of the Cel 1 assay by 4 weeks post-infection (Fig. 4a)." . . . _:b437200882 . _:b437201190 . _:b437200504 . _:b437201185 . _:b437201630 . _:b437200676 . _:b437200491 . _:b437200243 . _:b437201184 . . _:b437200894 . . _:b437201187 . . _:b437201186 . _:b437201197 . _:b437201406 . . . _:b437200783 . _:b437201196 . . _:b437201199 . _:b437201198 . _:b437201193 . _:b437201131 . . _:b437201192 . . _:b437201195 . _:b437201490 . _:b437201194 . _:b437201139 . _:b437201205 . . _:b437200516 . . _:b437201204 . . _:b437201879 . _:b437201207 . _:b8124668 . . . _:b437201206 . _:b437201524 . _:b437200577 . _:b437201201 . _:b437200718 . _:b437201200 . _:b437201008 . _:b437201203 . . _:b437200926 . _:b437201202 . . . _:b437201213 . _:b437200879 . _:b437200769 . _:b437201212 . _:b437200967 . _:b437201215 . _:b437201655 . _:b437200256 . _:b437201272 . _:b437201214 . _:b8124661 . _:b437201209 . . _:b437201208 . _:b437200379 . _:b437201211 . _:b437201210 . _:b437201478 . _:b437201221 . _:b437201220 . _:b437200628 . _:b437201223 . . _:b437201222 . _:b437201217 . _:b437201216 . _:b437201219 . _:b437200751 . _:b437201218 . _:b437200665 . . _:b437201229 . _:b437201969 . _:b437201228 . _:b437201498 . . . _:b437201231 . . _:b437201725 . _:b437201230 . _:b437201225 . _:b437200931 . _:b437201224 . _:b437201940 . _:b437201227 . _:b437201310 . _:b437201226 . _:b437201237 . . _:b437201236 . _:b437201849 . _:b437201239 . . _:b437201116 . . . _:b437201238 . _:b437201233 . _:b437201497 . _:b8124655 "CCR5-targeted ZFNs are currently being evaluated in a clinical trial targeting mature CD4+ T cells (>>19<<). However, disruption of CCR5 in HSC is likely to provide a more effective and long-term anti-viral effect, giving rise to CCR5-negative cells in both the lymphoid and myeloid compartments that HIV-1 infects." . . _:b437201809 . _:b437201232 . _:b437201235 . _:b437201234 . _:b437201671 . _:b437201245 . _:b437200329 . _:b437200834 . _:b437201244 . . _:b437200340 . . _:b437201247 . _:b437201344 . . _:b437201126 . _:b437201246 . _:b437200503 . _:b437201241 . _:b437200948 . . _:b437201031 . _:b437201240 . _:b437201066 . _:b437201789 . _:b437201243 . . . . _:b437201242 . . _:b437201253 . _:b437201252 . _:b437201255 . . . . _:b437201254 . _:b437201249 . _:b437201248 . . _:b437201029 . _:b437201251 . _:b437201250 . . . _:b437201261 . . _:b437201260 . . _:b437201600 . _:b437201263 . . _:b437201262 . . _:b437201466 . _:b437200476 . _:b437201221 . _:b437201257 . . _:b437201123 . _:b437201256 . _:b8124716 . _:b437201259 . . _:b437201258 . _:b437201269 . _:b437201612 . . . _:b437201268 . . . _:b437201271 . . _:b8124662 "Although non-viral methods such as nucleofection are attractive, this procedure has previously been associated with relatively high rates of toxicity in human CD34+ HSC and loss of engraftment potential (>>22<<), although more recently, less toxic outcomes have been described (23\u201325)." . . _:b437201270 . . _:b437201265 . . _:b437200979 . _:b437201264 . _:b437201224 . _:b437200255 . _:b437201445 . . _:b437201267 . _:b437200220 . _:b437201158 . _:b437201266 . . _:b437201277 . . . _:b437201276 . _:b437201279 . _:b437201848 . . _:b437201278 . _:b437201953 . _:b437201714 . . _:b437201273 . . _:b437200937 . _:b437200501 . _:b437201972 . _:b437200693 . _:b437201272 . . _:b437201275 . _:b437200569 . _:b437201274 . _:b437200484 . . _:b437200611 . _:b437201285 . _:b437201177 . _:b437201284 . . _:b437200615 . . _:b437200465 . . _:b437201287 . _:b437201642 . _:b437201286 . _:b437201281 . _:b437201280 . _:b437201283 . _:b437201282 . _:b8124705 . _:b437201293 . . _:b437201292 . . _:b437201904 . _:b437200918 . . _:b437201464 . _:b437201295 . _:b437201294 . _:b437201289 . . _:b437201288 . _:b437200610 . _:b437201291 . . _:b437200448 . _:b437201290 . _:b437201129 . _:b437201340 . _:b437201301 . . _:b437200566 . . _:b437201300 . _:b437201559 . . _:b437200965 . _:b437200861 . _:b437201303 . _:b437201286 . _:b8124679 . _:b8124653 . _:b437201302 . _:b437201814 . _:b437201297 . . _:b437200731 . _:b8124646 "(HSC) to generate HIV-resistant progeny that can suppress HIV-1 replication in vivo was demonstrated in an HIV-infected patient undergoing HSC transplantation from a homozygous CCR5\u039432 donor during treatment for acute myeloid leukemia (>>9<<). The donor cells were able to confer long-term control of HIV-1 replication and to restore the patient\u2019s CD4+ T cell levels in the absence of any antiretroviral drugs." . _:b437201296 . _:b437201299 . . _:b437201298 . _:b8124674 . _:b437201309 . _:b8124673 . . . . _:b437201308 . _:b437200242 . _:b437201311 . . _:b437201310 . . _:b437201305 . _:b437200840 . . _:b437201304 . _:b437201307 . . _:b437200745 . _:b437201306 . _:b437201317 . . . . _:b437201506 . _:b437201316 . . . . _:b437201319 . _:b437201288 . _:b437201318 . . . . _:b437201313 . _:b437201037 . . _:b437201676 . _:b437201312 . _:b437200991 . . _:b437201315 . . _:b437201314 . . _:b437201325 . . _:b437200348 . _:b437201324 . _:b437200225 . . _:b437201327 . _:b437201326 . _:b437201321 . _:b437201320 . _:b437201230 . _:b437201762 . _:b437200714 . _:b437201323 . _:b437201521 . _:b437201322 . _:b8124670 "Additionally, the animals did not experience any obvious toxicity or ill health, as has been reported for animals receiving higher radiation doses (>>27<<). Using this optimized engraftment procedure, we found that ZFN-treated HSC engrafted NSG mice as efficiently as untreated HSC controls (Fig. 1b), with no statistically significant difference between the two groups (Student\u2019s t test," . _:b437200657 . . _:b437201333 . _:b8124661 "Since DSB formation requires only transient expression of ZFNs, gene delivery methods that are suitable to express ZFNs include plasmid DNA nucleofection, integrase-defective lentiviral vectors and adenoviral vectors (16,19,>>21<<). Although non-viral methods such as nucleofection are attractive, this procedure has previously been associated with relatively high rates of toxicity in human CD34+ HSC and loss of engraftment potential (22), although more recently," . . _:b437201332 . _:b8124667 "any adverse effects on cell viability resulting from the nucleofection procedure may be offset by the high levels of CCR5 disruption achieved, as well as the speed and simplicity of the procedure when compared to viral vector systems (>>19<<,21)." . . _:b8124722 "patients treated with CCR5 inhibitors unless pre-existing CXCR4-tropic viruses were present before therapy, and resistance to the drugs can occur, instead, because of viral adaptations to the drug-bound form of the CCR5 co-receptor (66,>>67<<). Interestingly, the patient receiving the CCR5\u039432 HSC transplantation harbored CXCR4-utilizing viruses before the procedure, yet he did not experience a tropism switch, and instead, successfully controlled his HIV-1 infection long-term" . _:b437201451 . . . . _:b437201335 . _:b437201334 . _:b437201329 . . _:b437201873 . . _:b437201328 . . . _:b437201331 . _:b437201330 . _:b437200424 . _:b437200560 . _:b437201863 . _:b437201341 . . _:b437201340 . . _:b437201343 . _:b437201342 . _:b437201534 . _:b437201337 . . . _:b437200259 . _:b437201336 . _:b437200889 . _:b437200767 . _:b437201339 . . _:b437201761 . _:b437201338 . . _:b437201349 . _:b437200187 . _:b437201348 . _:b437201351 . _:b437201105 . _:b437200307 . _:b437201313 . _:b437200529 . _:b437201350 . . _:b437201551 . _:b437201345 . _:b437201284 . _:b437201344 . . _:b437201347 . . _:b437201388 . _:b437201763 . _:b437201346 . . _:b437201968 . . _:b8124718 . _:b437201357 . _:b437200540 . _:b437202001 . _:b437201798 . _:b437201356 . . _:b437201359 . _:b437201358 . _:b437201353 . _:b437200415 . . . _:b437200191 . _:b437201352 . _:b437200763 . . _:b437200279 . _:b437201355 . _:b437201354 . . _:b437201365 . _:b437201364 . _:b437201947 . . _:b437201367 . _:b437201366 . _:b437201896 . . _:b437201361 . . _:b8124703 . _:b437200639 . _:b437201360 . _:b437201363 . . . . _:b437201362 . _:b8124703 . . _:b437200440 . _:b437201373 . . _:b437201372 . _:b437201059 . _:b437201375 . . . . _:b437201996 . _:b437201374 . _:b437201479 . . . _:b437201369 . . _:b437201368 . . _:b437201371 . _:b437200433 . _:b437201370 . . _:b437202008 . _:b437201381 . _:b437201380 . _:b437201163 . . _:b437201383 . _:b437201390 . _:b437201382 . . . _:b437201377 . _:b437201422 . _:b437201376 . _:b437202007 . _:b437201186 . _:b437201114 . _:b437201379 . _:b437201378 . _:b437201908 . _:b437200815 . . _:b437201389 . _:b437201657 . _:b437201388 . _:b437201391 . _:b8124709 . _:b437201390 . _:b437201039 . _:b437201777 . _:b437201385 . _:b437201384 . . _:b437201977 . _:b437201387 . . _:b437201386 . _:b437201397 . . _:b8124712 . _:b437201396 . _:b8124694 . _:b437201399 . _:b437201088 . _:b437201398 . . _:b437201865 . _:b437201393 . _:b437201902 . _:b437201392 . . _:b437201380 . _:b437201395 . _:b437201997 . _:b437200245 . _:b437200198 . _:b437200271 . _:b437201394 . . _:b437201361 . _:b8124709 "recent finding that an AIDS leukemia patient undergoing HSC transplantation from a CCR5-negative donor was effectively cured of his infection, with undetectable levels of HIV-1 and rebounds in CD4 T cell levels despite discontinuing ART (>>9<<). As shown by our data, ZFN-modified HSCs retain full functionality and give rise to CCR5-negative cells in lineages relevant to HIV-1 pathogenesis, such as T cells and monocyte/macrophages." . _:b437201019 . _:b437201793 . _:b437201405 . _:b8124723 . _:b437201404 . . . _:b437201407 . _:b437200367 . . _:b437201406 . _:b437201401 . . . _:b437201400 . _:b437200286 . _:b437201403 . _:b437201591 . _:b8124729 . _:b437201402 . . . . _:b437200901 . . . _:b437200900 . . . _:b437200903 . _:b437201933 . _:b437200902 . _:b437201471 . . _:b437200897 . . . . _:b437200896 . _:b437200899 . _:b437200898 . _:b437200436 . . _:b437200909 . _:b437200489 . . _:b437200386 . . _:b437200908 . _:b437200311 . _:b437200911 . . _:b437200984 . _:b437200910 . _:b437201155 . _:b437200680 . _:b437201611 . _:b437200798 . _:b437200905 . . _:b437201842 . _:b437200709 . _:b437201452 . _:b437200904 . . . . _:b437200227 . . . . _:b437200907 . . _:b437200906 . _:b437200917 . . . _:b437200916 . . _:b437201743 . _:b437201529 . _:b437201945 . _:b437200919 . _:b437200918 . _:b437200913 . . _:b437200912 . . _:b437201838 . _:b437200586 . _:b437200915 . . . _:b437200914 . . _:b437200925 . _:b437200479 . _:b437200964 . _:b437200924 . _:b437201488 . . _:b437200927 . _:b8124699 . _:b437200926 . . _:b437201100 . _:b437200921 . . _:b437200920 . _:b437201586 . _:b437201564 . _:b8124728 "strategy, as recently described for HIV-infected individuals receiving autologous HSC engineered with anti-HIV ribozymes, where gene marked progeny were found at higher levels in patients who underwent treatment interruptions (>>70<<)." . _:b437200923 . _:b437200922 . _:b437201416 . _:b437200933 . . _:b437200932 . _:b437200935 . _:b437200934 . _:b437201259 . _:b437201353 . _:b437200929 . _:b437200928 . . _:b437201523 . . _:b437200931 . _:b437201826 . . _:b437200930 . _:b437200739 . _:b437201005 . _:b437200941 . _:b8124714 "Of note, certain HIV-infected patient populations, such as AIDS lymphoma patients, already undergo full ablation and autologous HSC rescue as part of their therapy (>>58<<), and thereby present an opportunity for HSC-based gene therapies (44)." . _:b437200940 . _:b437200943 . _:b437200942 . _:b437200937 . _:b437201880 . _:b437201707 . _:b437200936 . _:b8124649 "The targeted cell populations include both mature peripheral T cells or HSC, and the loss of CCR5 in HSC appears to have no adverse effects on hematopoiesis (>>12<<,13,15)." . . . _:b437200939 . _:b437201722 . _:b437201946 . _:b437200938 . _:b437201212 . _:b437200369 . . _:b437200949 . . _:b437200948 . . . _:b437200951 . . _:b437200260 . . _:b437200950 . . . _:b437200945 . _:b437200944 . _:b437200462 . _:b437201702 . . _:b437201590 . _:b437200947 . _:b437200737 . . _:b437200946 . _:b437201631 . _:b437201824 . _:b8124702 "ZFNs can be designed to bind to a specific genomic DNA sequence and, by generating a targeted DSB, can affect the efficient, specific and permanent knockout of the targeted gene (>>19<<,45\u201347). In contrast to other genetic approaches to the control of HIV-1, ZFN-mediated gene disruption is a one-time, \u201Chit-and-run\u201D process that requires only transient expression of the ZFNs in HSC during a brief period of ex vivo culture." . _:b437200957 . _:b437201481 . _:b437200956 . . . _:b437200959 . _:b437201716 . _:b437201189 . _:b437200958 . _:b437200953 . _:b437201343 . _:b437200952 . _:b437200955 . . _:b437200954 . . . _:b437201886 . . _:b437200965 . _:b437200552 . _:b437200964 . _:b437200967 . _:b437200925 . _:b437200966 . . _:b437201917 . _:b437200961 . . _:b437200570 . _:b437200960 . _:b437200963 . . _:b437201324 . . _:b437200962 . _:b437200973 . _:b437200746 . _:b437200972 . _:b437201862 . _:b437200975 . . _:b8124715 . _:b437200974 . . _:b437200969 . _:b437200968 . _:b437200499 . _:b437200971 . _:b437200495 . . _:b437200970 . _:b437200981 . _:b437200701 . _:b437200980 . _:b8124704 . _:b437200983 . _:b8124727 . _:b437201375 . _:b437201975 . _:b437201992 . . _:b437200982 . . . _:b437200977 . _:b437200976 . . _:b437200979 . . _:b437201677 . . _:b437200978 . _:b437200642 . _:b437201720 . . _:b437200989 . _:b437200383 . _:b437201840 . _:b437202002 . _:b437200988 . _:b8124673 "This strain of HIV-1 has previously been used in humanized mouse models, where it causes a robust infection and significant CD4+ T cell depletion (35,>>36<<). These observations mimic the human infection, where depletion of CD4+ CCR5+ lymphocytes results from a combination of direct infection, systemic immune activation and the upregulation of CCR5 on thymocytes (36,37,38). Following" . _:b437201954 . _:b437200991 . . _:b437200719 . _:b437200736 . _:b437200990 . . _:b437200568 . . _:b437200408 . _:b437200985 . . . _:b437200984 . _:b437200987 . _:b437201914 . . _:b437200986 . . _:b437201117 . _:b437200514 . _:b437200997 . . _:b437200996 . _:b437200999 . . _:b437201241 . _:b437200998 . _:b437201431 . _:b437200607 . _:b437200993 . _:b437200667 . _:b437200518 . _:b437200992 . _:b437201112 . _:b437201132 . _:b437200995 . . _:b437200994 . _:b437201347 . _:b437201005 . _:b437201414 . _:b437201004 . _:b437200795 . _:b437201732 . . _:b437201007 . _:b437200907 . _:b437201006 . . _:b437201001 . _:b437201000 . . _:b437200523 . _:b437201003 . _:b437201148 . _:b437201002 . . _:b437201013 . . _:b437201012 . . _:b437201015 . . _:b437201750 . . _:b437201740 . . _:b437201014 . . . . _:b437200982 . _:b437201009 . . _:b437201008 . _:b437200841 . . _:b437201011 . _:b437201010 . _:b437201021 . _:b437201020 . _:b437200320 . _:b437201023 . _:b8124691 "extracting RNA from 5 \u00D7 105 cells using the master pure complete DNA and RNA purification kit (Epicentre Biotechnologies) and performing Taqman qPCR using a primer and probe set targeting the HIV-1 LTR region, as previously described (>>72<<). In addition, p24 levels were measured in blood samples by ELISA." . _:b437201021 . _:b437201022 . _:b437201832 . . . _:b437201017 . _:b437201166 . _:b437201016 . . _:b437201042 . _:b437201019 . _:b437201878 . _:b437201748 . _:b437201018 . _:b437201819 . _:b437201029 . . _:b437201171 . _:b437201028 . _:b8124700 "In this regard, the CCR5 co-receptor is a particularly attractive target because of the HIV-resistant phenotype of homozygous CCR5\u039432 individuals (>>3<<), who tolerate this genotype with minimal consequences (5\u20137)." . . _:b437201658 . _:b437201031 . . _:b437201030 . _:b437200673 . _:b437201025 . _:b8124712 . _:b437200616 . _:b437201024 . _:b437201180 . _:b437201027 . _:b437201279 . . . _:b437201026 . _:b437200385 . _:b437200732 . _:b437201037 . _:b437201036 . _:b437200201 . _:b437201855 . . _:b437201039 . _:b8124651 "The targeted cell populations include both mature peripheral T cells or HSC, and the loss of CCR5 in HSC appears to have no adverse effects on hematopoiesis (12,13,>>15<<)." . _:b437201038 . _:b437200855 . _:b437201839 . _:b437201033 . . _:b437202004 . . _:b437201032 . _:b437201435 . . _:b437201035 . _:b437201034 . . _:b8124701 . _:b437201556 . . _:b437201045 . _:b437201044 . . _:b437200766 . _:b437201047 . _:b437201321 . _:b437201046 . _:b437201538 . . _:b437201041 . . _:b437201623 . _:b437201041 . . _:b437201040 . _:b8124649 . . _:b437201043 . _:b8124648 . _:b437201042 . _:b437201053 . _:b8124697 . _:b437201436 . _:b437200606 . _:b437201052 . _:b437201898 . _:b437201360 . _:b437201055 . _:b437200833 . _:b437201054 . _:b437200910 . . _:b437201049 . . _:b437201048 . _:b437200239 . . _:b437201460 . _:b437201051 . _:b437201050 . _:b437201061 . . _:b437201060 . _:b437201474 . _:b437200909 . _:b437201693 . _:b437201063 . _:b437201528 . _:b437201370 . _:b437200597 . . . _:b437201062 . _:b437200224 . _:b437200196 . _:b437201057 . _:b437200371 . _:b437200197 . _:b437200198 . . _:b437200199 . . _:b437201056 . _:b437200192 . _:b437200193 . _:b437200194 . _:b437200195 . _:b437200204 . _:b437201059 . _:b437200205 . _:b437200206 . . _:b437200207 . _:b437200583 . _:b437200493 . _:b437201058 . _:b437200200 . _:b437200201 . _:b437200202 . _:b437200203 . _:b437200477 . _:b437200212 . _:b437201069 . _:b437200213 . _:b437200214 . _:b437200215 . _:b437200208 . _:b437201068 . . . _:b437200209 . . _:b437201394 . . _:b437200210 . _:b437200211 . . _:b437201071 . _:b437200220 . _:b437200221 . _:b437200222 . _:b437200223 . . _:b437200216 . _:b437201070 . . _:b437200217 . _:b437200218 . _:b437200219 . _:b437200636 . _:b437200228 . _:b437201065 . _:b437200229 . _:b437200230 . _:b437200231 . _:b437200224 . _:b437201064 . _:b437200225 . . _:b437200226 . _:b437200227 . _:b437200236 . _:b437201067 . _:b437200237 . _:b437201650 . _:b437200238 . _:b437200239 . _:b437200580 . _:b437200232 . _:b437201066 . _:b437200233 . _:b437201244 . _:b437201045 . _:b437200234 . _:b437200235 . _:b437200244 . _:b437201077 . _:b437200245 . _:b437200246 . . _:b437200247 . _:b437200240 . _:b437201076 . _:b437200241 . _:b437200242 . _:b437201441 . _:b437200243 . _:b437200252 . _:b437201079 . . _:b437200253 . . _:b437200254 . _:b437200255 . . _:b437200195 . _:b437201078 . _:b437200248 . _:b437200249 . _:b437200250 . _:b437200251 . _:b437200687 . . _:b437201073 . . . _:b437201965 . _:b437201072 . . . _:b437200625 . _:b437201891 . _:b437201075 . . _:b437201074 . . _:b437201085 . . _:b437201084 . . _:b437201087 . . _:b8124671 . _:b437201086 . _:b437201263 . . _:b437201081 . . _:b437201136 . _:b437201080 . _:b437201656 . _:b437201083 . _:b437201193 . . _:b437201004 . _:b437201082 . _:b437200715 . . _:b437201093 . _:b437200651 . _:b437200753 . _:b437201092 . _:b437201618 . . _:b437201095 . _:b437200188 . . . . _:b437200189 . _:b437200190 . . _:b437200191 . _:b437201094 . _:b437200186 . _:b437200187 . _:b437201104 . _:b437200324 . _:b437201089 . _:b437200325 . _:b437200326 . _:b437200327 . _:b437201476 . _:b437200320 . _:b437201088 . _:b437200321 . _:b437200322 . _:b437200323 . _:b437200332 . _:b437201091 . _:b437200333 . _:b437200334 . _:b437200335 . _:b437200328 . _:b437201090 . _:b437200329 . . _:b437200330 . _:b437200331 . _:b437200340 . _:b437201101 . _:b437200341 . _:b437201194 . _:b437200342 . _:b437200343 . _:b437200336 . _:b437201100 . _:b437200337 . _:b437200338 . _:b437200339 . . _:b437201103 . _:b437200348 . _:b437200349 . _:b437200350 . _:b437200351 . _:b437200344 . _:b437201102 . _:b437200397 . _:b437200345 . _:b437200346 . _:b437201581 . _:b437200347 . _:b437200356 . _:b437201097 . _:b437200357 . _:b437200358 . _:b437201737 . _:b437200359 . _:b437200352 . _:b437201096 . _:b437200353 . _:b437200354 . _:b8124720 . _:b437200218 . _:b437200355 . _:b437200364 . _:b437201099 . _:b437200365 . _:b437200366 . _:b437200367 . _:b437200360 . _:b437201098 . _:b437200361 . _:b437200362 . _:b437200363 . _:b437200372 . _:b437201109 . _:b437200373 . _:b437200374 . . _:b437200375 . _:b437200467 . _:b437201108 . . _:b437200368 . _:b437200369 . _:b437201859 . _:b437200370 . _:b437200371 . _:b437201504 . _:b437200954 . _:b437200380 . _:b437201111 . _:b437200381 . . _:b437200382 . _:b437200383 . _:b437200376 . _:b437201110 . _:b437200377 . _:b437200378 . _:b437201566 . _:b437200379 . _:b437200260 . _:b437201105 . _:b437200261 . _:b437201778 . _:b437200430 . _:b437200262 . _:b437200263 . _:b437200256 . _:b437201104 . _:b437200257 . _:b437200258 . _:b437201922 . _:b437200259 . _:b437200268 . _:b437201448 . _:b437201107 . _:b437200269 . _:b437200270 . _:b437200271 . _:b437200264 . _:b437201106 . _:b437200265 . _:b437200419 . _:b437201941 . _:b437200266 . _:b437200768 . _:b437200267 . _:b437200276 . _:b437201117 . _:b437200277 . _:b437200278 . _:b437200279 . _:b437200272 . _:b437201116 . _:b437200273 . _:b437200274 . _:b437200275 . _:b437200284 . _:b437201119 . . _:b437200285 . _:b437200286 . _:b437200813 . _:b437200287 . _:b437200234 . _:b437200280 . _:b437201118 . _:b437200281 . . _:b437200282 . _:b437200283 . _:b437201627 . _:b437200292 . _:b437201220 . _:b437201113 . _:b437200293 . _:b437200588 . _:b437200294 . _:b437200617 . _:b437200295 . _:b437200288 . _:b437201112 . . _:b437200289 . _:b437200290 . _:b437200291 . _:b437200300 . _:b437201115 . . _:b437200301 . _:b437200302 . _:b437200303 . _:b437200296 . _:b437201114 . . _:b437200297 . _:b437200298 . _:b437200299 . _:b437200308 . _:b437201125 . _:b437200309 . _:b437200310 . _:b437200311 . _:b437200304 . _:b437201124 . . _:b437200305 . _:b437200306 . _:b437200307 . _:b437201760 . _:b437201065 . _:b437200316 . _:b437201127 . _:b437200317 . _:b437200915 . _:b437201202 . _:b437200318 . _:b437200319 . _:b437200312 . _:b437201126 . . _:b437200313 . _:b437200314 . _:b437201376 . _:b437200315 . _:b437201121 . . _:b437201120 . . _:b437201123 . _:b437201122 . _:b437201505 . . _:b437201133 . . _:b437200727 . _:b437201014 . _:b437201132 . . _:b437201386 . _:b437201135 . _:b437200928 . . _:b437200972 . . _:b437201134 . . _:b437201129 . _:b437200640 . . _:b437201128 . _:b437201856 . _:b437201897 . _:b437201131 . _:b437201130 . _:b437200206 . _:b437201141 . _:b437201532 . _:b437201140 . _:b8124665 "Similar results were also achieved using CD34+ cells isolated from fetal liver (data not shown). Previous studies in both cell lines (>>16<<) and primary human T cells (19) have shown that the percentage of bi-allelically modified cells in a population treated with ZFNs is between 30\u201340% of the total number of disrupted alleles detected by the Cel I assay, so that we estimate" . . _:b437201143 . _:b437201142 . _:b437201137 . _:b437201136 . _:b437201139 . _:b437200618 . _:b437201138 . _:b437200398 . _:b437201149 . _:b437201812 . . _:b437201148 . . _:b437201151 . . _:b437201150 . _:b437200694 . _:b437201145 . _:b437201144 . _:b437200210 . _:b437200589 . _:b437201147 . _:b437201146 . . _:b437201606 . _:b437200685 . . . _:b437200434 . . . . _:b437201738 . _:b437200290 . . _:b437200986 . . . _:b437200524 . . _:b437201537 . . . _:b437201890 . _:b437200790 . . _:b437200946 . _:b437201182 . _:b437201090 . _:b437201718 . _:b437200755 . . . . . _:b437200531 . _:b437200893 . . _:b437201152 . _:b437200575 . . . _:b437200722 . _:b437201948 . . . _:b8124712 "This is especially true in the GALT, which represents a key battleground in HIV-1 infection (>>54<<\u201356). We also noted a strong selective pressure for CCR5-negative cells at the level of the thymus. This suggests that T cells would be selected to be CCR5-negative at both a precursor stage in the thymus, as well as the effector CD4+" . . . . _:b437201063 . _:b437200735 . _:b437200323 . _:b437200273 . _:b437201418 . _:b437200708 . _:b437200709 . _:b437200710 . _:b437200711 . _:b437200704 . _:b437200705 . _:b437200706 . _:b437200707 . _:b437200716 . _:b437200717 . _:b437200718 . _:b437200719 . _:b8124726 . _:b437200712 . _:b437200713 . _:b437201704 . . _:b437200714 . _:b437200715 . _:b437200724 . _:b437200725 . _:b437200726 . _:b437200727 . _:b437200720 . _:b437200721 . _:b437200722 . _:b437200723 . _:b437200732 . _:b437200733 . _:b437200734 . _:b437200735 . _:b437200728 . _:b437200729 . _:b437200730 . . _:b437200731 . _:b437200740 . _:b437200741 . _:b437200742 . _:b8124716 . _:b437200743 . _:b437200736 . _:b437200737 . _:b437200738 . _:b437200739 . _:b437200748 . _:b437201569 . _:b437201290 . _:b437200749 . _:b437201934 . _:b437200750 . _:b437200751 . _:b437201299 . _:b437200744 . _:b437200251 . _:b437200745 . . _:b437200746 . _:b437200747 . _:b437200756 . _:b437201511 . _:b437200757 . _:b437200758 . _:b437200759 . _:b437200752 . _:b437200753 . _:b437200754 . _:b437200755 . _:b437200764 . _:b437200765 . _:b437200766 . _:b437200767 . _:b437200760 . _:b437200761 . _:b437200762 . _:b437200763 . _:b437200644 . _:b8124688 "disrupted by ZFN treatment was measured by performing PCR across the ZFN target site followed by digestion with the Surveyor (Cel 1) nuclease (Transgenomic, Omaha, NE), which detects heteroduplex formation, as previously described (>>19<<). Briefly, genomic DNA was extracted from mouse tissues and subject to nested PCR amplification using human CCR5 specific primers, with the resulting radiolabeled products digested with Cel I nuclease and resolved by PAGE." . _:b437200645 . _:b437200646 . _:b437200647 . _:b437200640 . _:b437200641 . _:b437200642 . _:b437201159 . _:b437200643 . _:b437200652 . _:b437200653 . _:b437200654 . _:b437200655 . _:b437200648 . _:b8124695 "Immunohistochemical analysis of human CD3 and CD4 expression, respectively, in the small intestine and spleen tissue from HSC engrafted mice was performed on fixed paraffin embedded tissue sections, as previously described (>>73<<,75). Controls included isotype matched non-specific antibodies, and unengrafted NSG mice." . _:b437200649 . _:b8124719 . _:b437200650 . _:b437200651 . _:b437200660 . _:b437200661 . _:b437200662 . . _:b437200663 . _:b437200442 . _:b437200656 . _:b437200657 . _:b437200658 . _:b437200659 . _:b437200668 . _:b437200669 . _:b437200670 . _:b437200671 . . . _:b437200664 . _:b437200665 . _:b437200666 . . _:b437200667 . _:b437200647 . . _:b437200676 . . _:b437200677 . _:b437200678 . _:b437201801 . _:b437200679 . _:b437200672 . _:b437200673 . . _:b437200674 . _:b437200546 . _:b437200675 . _:b437200684 . _:b437200685 . _:b437200686 . _:b437200687 . _:b437200405 . _:b437200680 . _:b437200681 . _:b437200682 . _:b437200683 . _:b437200674 . _:b437200692 . _:b437200444 . _:b437200693 . _:b437200694 . _:b437200695 . . . _:b437200688 . . _:b437200312 . _:b8124665 . _:b437200689 . _:b437200690 . _:b437201185 . _:b437200691 . _:b437200700 . _:b437200701 . _:b8124676 "These observations mimic the human infection, where depletion of CD4+ CCR5+ lymphocytes results from a combination of direct infection, systemic immune activation and the upregulation of CCR5 on thymocytes (36,37,>>38<<). Following infection, blood samples were collected from the mice every two weeks and analyzed for HIV-1 RNA levels, T cell subsets and the extent of CCR5 disruption. At 8\u201312 weeks post-infection, animals were euthanized and necropsy" . _:b437201570 . _:b437200702 . . _:b437200703 . _:b437200696 . _:b437200697 . _:b437200698 . _:b437200699 . _:b437200836 . _:b437200837 . _:b437200827 . _:b437201768 . _:b437200838 . _:b437200839 . _:b437200832 . _:b437200833 . _:b437200834 . _:b437200835 . _:b437200844 . _:b437200845 . _:b437200846 . _:b437201160 . _:b437200407 . _:b437200847 . _:b437200840 . _:b437200841 . _:b8124659 . _:b437200842 . _:b437201450 . _:b437200843 . _:b437200852 . _:b437200853 . _:b437200854 . _:b437201339 . _:b437200231 . _:b437200855 . _:b437200848 . _:b437200849 . _:b437200850 . _:b437200851 . _:b437200860 . _:b437200861 . _:b437201477 . _:b437200862 . . _:b437200863 . _:b437200856 . _:b437200857 . _:b437200858 . _:b437200859 . _:b437200868 . _:b437200594 . _:b437200869 . . _:b437200870 . _:b437200871 . . _:b437200864 . _:b437200865 . _:b437200866 . _:b437200395 . _:b437200867 . _:b437200876 . _:b437200877 . _:b437200878 . _:b437200879 . _:b437201888 . _:b437200872 . _:b8124686 "One \u00D7 106 cells were nucleofected with 2.5 \u03BCg each of a plasmid pair expressing ZFNs binding upstream (ZFN-L) or downstream (ZFN-R) of codon Leu55 within TM1 of human CCR5 (>>19<<). The CD34+ cell/DNA mix was processed in an \u00D7 series Amaxa Nucleofector (Lonza) using the U-01 setting and the human CD34+ nucleofector solution, according to the manufacturer\u2019s instructions. Following nucleofection, cells were" . _:b437200873 . _:b437200874 . _:b437201684 . _:b437200875 . _:b437200884 . _:b437200885 . _:b437200886 . _:b437200887 . _:b437200880 . _:b437200881 . . _:b437200882 . . _:b437200883 . . _:b437200892 . . _:b437200893 . . _:b437200894 . _:b437200895 . . _:b437200474 . _:b437200888 . . _:b437200889 . _:b437200890 . _:b437200891 . _:b437200772 . . _:b8124652 . _:b437200773 . _:b437200774 . . _:b437200775 . _:b437200768 . _:b437200728 . _:b437200769 . _:b437200770 . _:b437200771 . _:b437200780 . _:b437200781 . _:b437200782 . _:b437200783 . _:b437201937 . . _:b437200776 . _:b437200777 . _:b437200778 . _:b437200779 . _:b437200298 . _:b437200788 . . . _:b437200789 . _:b437200790 . _:b437201401 . _:b437200791 . _:b437201828 . . _:b437200784 . _:b437200785 . _:b437200786 . . _:b437200787 . _:b437200796 . _:b437200797 . _:b437200798 . _:b8124677 "Changes in the ratio of CD4+ to CD8+ T cells in the peripheral blood are characteristic of progressive infection in AIDS patients (>>39<<,40). We therefore examined the CD4:CD8 ratio in the peripheral blood samples from individual mice, both pre- and post-infection." . _:b437200799 . _:b437201399 . . _:b437200792 . _:b437200793 . . _:b437200794 . _:b437200795 . _:b437200804 . _:b437200805 . _:b437200806 . _:b437200807 . _:b437201744 . _:b437200800 . _:b437200801 . _:b437200802 . _:b437200803 . _:b437200812 . _:b437200813 . _:b437200814 . _:b437201033 . _:b437200815 . _:b437200808 . _:b437200809 . _:b437201118 . _:b437200810 . _:b437200811 . . _:b437200820 . . _:b437200821 . _:b437200502 . _:b437200822 . _:b437200823 . _:b437200816 . . _:b437200817 . _:b437200818 . _:b437200819 . _:b437200828 . _:b437201040 . . _:b437200829 . _:b437200830 . _:b437200831 . _:b437200824 . _:b437200825 . . _:b437200826 . . _:b437200827 . . _:b437200452 . _:b437200453 . _:b437200454 . _:b437201176 . _:b437200455 . _:b437200448 . _:b437200449 . _:b437200450 . _:b437201273 . . _:b437200451 . _:b437200460 . _:b437200461 . _:b437200462 . _:b437200463 . _:b437200456 . _:b8124693 "Intestinal samples were processed as previously described (>>73<<), with the modification that the mononuclear cell population was isolated following incubation in citrate buffer and collagenase enzyme for 2 hrs, followed by nylon wool filtration (Amersham Biosciences, Sweden) and ficoll-hypaque" . _:b437200457 . _:b437200458 . . _:b437200459 . _:b437200468 . _:b437201632 . _:b437200469 . _:b437200470 . _:b437200471 . _:b437200464 . . _:b437201833 . _:b437200465 . _:b437200466 . _:b437200467 . _:b437200476 . . _:b437200477 . _:b437200478 . _:b437200479 . _:b437200472 . _:b437200473 . _:b437200474 . _:b437201147 . _:b437200475 . _:b437200484 . _:b437200485 . _:b437200486 . _:b437200487 . _:b437200480 . . _:b437200481 . _:b437200482 . . _:b437201649 . _:b437200483 . . _:b437200209 . _:b437200492 . . _:b437200193 . _:b437200493 . . _:b437200494 . _:b437200319 . _:b437200495 . . _:b437200488 . _:b437200489 . _:b437200490 . _:b437200491 . _:b437200500 . _:b437200501 . _:b437200502 . . _:b437200503 . _:b437200496 . . _:b437200497 . _:b437200498 . _:b437200230 . _:b437200499 . _:b437201619 . _:b437200508 . _:b437200509 . _:b437201178 . _:b437200510 . . _:b437200511 . . _:b437200504 . . . _:b437200505 . _:b437200506 . _:b437200507 . . _:b437200388 . _:b437200389 . _:b437201396 . _:b437200530 . _:b437200390 . _:b437200906 . _:b437200391 . _:b437201930 . _:b437200384 . _:b437201377 . . _:b437200385 . _:b437200386 . _:b437200387 . _:b437200396 . . _:b437200397 . _:b437200471 . . _:b437200398 . _:b437200399 . _:b437200392 . _:b437200393 . _:b437200394 . _:b437200395 . _:b437200404 . . _:b437200405 . _:b437200406 . _:b437200407 . _:b437200400 . _:b437200401 . _:b437200402 . _:b437200403 . _:b437200500 . _:b437200412 . . _:b437201379 . _:b437200413 . _:b8124658 _:b8124684 . _:b437200414 . _:b437201161 . _:b437200415 . _:b437200408 . _:b8124658 _:b8124680 . _:b437200409 . _:b8124658 _:b8124681 . _:b437200410 . _:b8124658 _:b8124682 . _:b437200411 . _:b8124658 _:b8124683 . _:b437200420 . _:b8124658 _:b8124676 . _:b437200421 . _:b8124658 _:b8124677 . _:b437200422 . _:b8124658 _:b8124678 . _:b437201926 . _:b437201648 . _:b437200423 . _:b8124658 _:b8124679 . _:b437200416 . . _:b437200417 . _:b8124658 _:b8124672 . _:b8124658 _:b8124673 . _:b437200418 . _:b8124658 _:b8124674 . _:b437200419 . _:b8124658 _:b8124675 . _:b437200428 . _:b437200429 . . _:b437200430 . _:b437200431 . _:b437201931 . _:b437200424 . . _:b437200425 . _:b437200426 . _:b437200427 . _:b437200436 . _:b437200437 . _:b437200438 . _:b437200439 . _:b437200305 . _:b437200432 . . _:b437200433 . _:b437200434 . _:b437201052 . _:b437200435 . _:b437201217 . _:b437200444 . _:b437201613 . _:b437200445 . _:b437200446 . _:b437201022 . _:b437200644 . _:b437200447 . _:b437200440 . _:b437200441 . _:b437200442 . . . _:b437200443 . _:b437200580 . _:b437200581 . _:b8124643 "A genetic mutation that deletes 32 bp from the CCR5 gene (CCR5\u039432) is relatively common in Western European populations and provides profound resistance to HIV-1 infection and AIDS in homozygotes (>>3<<,4). The absence of any other significant phenotype associated with a lack of CCR5 (5\u20137) has spurred the development of therapies aimed at blocking the virus-CCR5 interaction, and CCR5 antagonists have proved to be an effective salvage" . _:b437200582 . _:b437200583 . _:b437200944 . _:b437200376 . _:b437200576 . _:b437200577 . _:b437200578 . _:b437200579 . _:b437200720 . _:b437200588 . _:b437200589 . _:b437200590 . _:b437200591 . _:b437201518 . _:b437200584 . _:b437200453 . _:b437200585 . _:b437200586 . _:b437200587 . _:b437200596 . _:b437200597 . _:b437200598 . _:b437200599 . . _:b437200592 . _:b437200593 . _:b437200594 . _:b437200595 . _:b437200604 . _:b437200605 . . _:b437200606 . . _:b437200607 . _:b437200600 . _:b437200922 . . _:b437200601 . _:b437200602 . _:b437200603 . . _:b437201593 . _:b437200612 . _:b437200613 . . _:b437200614 . _:b437200615 . _:b437200608 . _:b437200609 . _:b437200610 . _:b437200611 . _:b437200620 . _:b437200621 . _:b437200622 . _:b437200623 . _:b437200616 . _:b437200617 . . _:b437200618 . _:b437200619 . . _:b437200628 . _:b437200629 . _:b437200630 . _:b437200631 . _:b437200624 . _:b437200625 . _:b437200626 . _:b437200627 . _:b437200636 . _:b437200416 . _:b437200637 . _:b437200638 . _:b437200639 . _:b437200632 . _:b437200633 . _:b437200634 . _:b437200635 . _:b437200516 . _:b437200517 . . _:b8124671 . _:b437200518 . _:b437200519 . _:b437200512 . _:b437200513 . _:b437200514 . _:b437200515 . _:b437200553 . _:b437200524 . . _:b437200525 . _:b437200526 . _:b437200527 . _:b437200520 . _:b437200521 . _:b437201893 . _:b437201754 . _:b437200522 . _:b437200523 . _:b437201986 . _:b437200935 . _:b437200532 . _:b437200533 . . _:b437201510 . _:b437200534 . . _:b437200535 . _:b437200528 . _:b437200327 . _:b437201351 . _:b437200529 . _:b437200530 . _:b437200531 . _:b437200540 . . _:b437201889 . _:b437200541 . _:b437200542 . _:b437200543 . _:b437200536 . _:b437200537 . _:b437200538 . _:b437200539 . _:b437200284 . _:b437200548 . _:b437200549 . _:b437200550 . _:b437200551 . _:b437200544 . _:b437201831 . _:b437200545 . _:b437200546 . _:b437200547 . . _:b437200556 . . _:b437200557 . . _:b437200558 . . _:b437200559 . _:b437200552 . _:b437200553 . _:b437200554 . _:b437200555 . _:b437200564 . _:b437201250 . . _:b437201944 . _:b437200641 . _:b437200565 . _:b437200566 . _:b437200554 . _:b437200567 . . _:b437200560 . _:b437200561 . _:b437201167 . _:b437200562 . . _:b437200563 . _:b437200572 . _:b437200573 . _:b437200574 . _:b437200575 . _:b437200568 . _:b437200569 . . _:b437201830 . _:b437200570 . _:b437200571 . _:b8124671 . . _:b437201547 . . . _:b437200726 . _:b437200678 . _:b437200426 . _:b437201709 . _:b8124699 "Consequently, an alternate approach to controlling HIV-1 replication that is being considered is the engineering of the body\u2019s immune cells to be resistant to infection by the virus (>>44<<). In this regard, the CCR5 co-receptor is a particularly attractive target because of the HIV-resistant phenotype of homozygous CCR5\u039432 individuals (3), who tolerate this genotype with minimal consequences (5\u20137). In the present study, we" . _:b437200431 . . . _:b437200924 . _:b437200752 . _:b437201278 . _:b437200747 . . _:b437200696 . . . . _:b437200993 . . . _:b437201653 . _:b437201932 . _:b437200517 . _:b437200331 . . . _:b437200883 . . . _:b437200544 . _:b437201064 . . . . . . _:b437200316 . . . . . _:b437200228 . . _:b437201820 . _:b437200361 . _:b437201629 . . _:b437201899 . _:b437201695 . _:b437201094 . . _:b437201200 . _:b437200482 . _:b437200675 . _:b437201588 . . _:b437200936 . _:b8124672 . _:b437201925 . _:b437201348 . . . . . . _:b437200985 . . _:b8124641 _:b8124642 . . _:b8124641 _:b8124643 . _:b8124641 _:b8124644 . _:b8124641 _:b8124645 . _:b8124641 _:b8124646 . _:b8124641 _:b8124647 . _:b8124641 _:b8124648 . _:b8124641 _:b8124649 . _:b8124641 _:b8124650 . _:b8124641 _:b8124651 . . _:b8124641 _:b8124652 . _:b8124641 _:b8124653 . _:b8124641 _:b8124654 . . _:b8124641 _:b8124655 . _:b437201058 . _:b437200267 . . _:b8124641 _:b8124656 . _:b8124641 _:b8124657 . _:b437201345 . _:b437201614 . _:b437201520 . . _:b437200244 . . . . . _:b8124679 "Depletion of both single and double positive thymocytes has been reported to occur as a consequence of the up-regulation of CCR5 on these cells following HIV-1 infection (>>36<<,38), and likely contributed to both the observed depletion of the thymus, as well as a reduction in the numbers of mature CD4+ and CD8+ T cells observed in other tissues." . . _:b437200562 . . _:b437200511 . _:b437201604 . _:b437201318 . _:b437200921 . . . . . _:b8124658 _:b8124668 . _:b8124658 _:b8124669 . _:b8124658 _:b8124670 . . _:b8124658 _:b8124671 . _:b8124658 _:b8124664 . _:b437200927 . _:b8124658 _:b8124665 . _:b437201967 . _:b8124658 _:b8124666 . _:b8124658 _:b8124667 . _:b8124658 _:b8124660 . _:b8124658 _:b8124661 . _:b8124658 _:b8124662 . _:b437201565 . . _:b8124658 _:b8124663 . . . _:b8124658 _:b8124659 . _:b437201049 . . . _:b437200555 . . . . . . . . . . _:b437200920 . _:b437200318 . _:b437201124 . . . _:b437200593 . _:b8124697 "Effective ART often involves costly, multi-drug regimens that are not well tolerated by a significant percentage of patients (>>42<<), and even successful adherence to ART does not eradicate the virus, so that a rapid rebound in HIV-1 levels can occur if therapy is discontinued (43)." . . _:b437201442 . . _:b437201637 . . _:b437201911 . _:b437200666 . . . _:b437201187 . . _:b437200468 . . . _:b437200602 . . _:b437200287 . _:b437201030 . _:b437201993 . _:b437201924 . _:b437200987 . . _:b437200545 . _:b437200887 . . . . . . . . . _:b437201942 . _:b437200390 "6"^^ . . _:b437201616 . _:b437200391 "6"^^ . _:b437201434 . . _:b437200388 "6"^^ . _:b437201172 . _:b437200389 "6"^^ . . _:b437200865 . _:b437200386 "6"^^ . _:b437201823 . _:b437200387 "6"^^ . . _:b437200963 . _:b437200772 . . _:b437200384 "6"^^ . _:b437200385 "6"^^ . _:b437200398 "6"^^ . . _:b437201682 . . _:b437200399 "6"^^ . _:b437200422 . . _:b437201723 . _:b437200396 "6"^^ . . _:b437200397 "6"^^ . _:b437201057 . . _:b437200760 . _:b437200394 "6"^^ . . _:b437200395 "6"^^ . . _:b437200392 "6"^^ . _:b437200393 "6"^^ . . . _:b437200406 "6"^^ . _:b437200407 "6"^^ . _:b437200404 "6"^^ . _:b437200405 "6"^^ . _:b437200402 "6"^^ . _:b437201107 . _:b437201567 . _:b437200403 "6"^^ . _:b437200400 "6"^^ . _:b437201726 . _:b437200401 "6"^^ . . _:b437200414 "6"^^ . . _:b437200541 . _:b437200415 "6"^^ . _:b437200248 . . _:b437200412 "6"^^ . _:b437200413 "6"^^ . . . _:b437200410 "6"^^ . _:b437201984 . _:b437201681 . _:b437200411 "6"^^ . _:b437200408 "6"^^ . _:b437200409 "6"^^ . _:b437201818 . . _:b437200579 . . _:b437200422 "6"^^ . _:b437200208 . _:b437201976 . _:b437200423 "6"^^ . . . _:b437200420 "6"^^ . _:b437201980 . . _:b437200421 "6"^^ . _:b8124648 "Towards this goal, various gene therapy approaches to block CCR5 expression are currently being evaluated, including the expression in hematopoietic cells of CCR5-specific intrabodies, ribozymes and siRNAs (>>11<<\u201314). The targeted cell populations include both mature peripheral T cells or HSC, and the loss of CCR5 in HSC appears to have no adverse effects on hematopoiesis (12,13,15)." . _:b437200418 "6"^^ . _:b437200419 "6"^^ . _:b437200859 . _:b437200416 "6"^^ . _:b437200417 "6"^^ . _:b437200873 . . . _:b437200430 "6"^^ . . . _:b437200269 . _:b437200428 "6"^^ . . _:b437201951 . _:b437200431 "5"^^ . _:b437200429 "6"^^ . . _:b437201091 . . _:b437201601 . _:b437200426 "6"^^ . _:b437200427 "6"^^ . _:b437200424 "6"^^ . _:b437200425 "6"^^ . _:b437200650 . _:b437200778 . _:b437200437 "5"^^ . . _:b437200853 . _:b437201651 . _:b437200436 "5"^^ . . _:b437200439 "5"^^ . _:b437200455 . _:b437200438 "5"^^ . _:b437201786 . _:b437200433 "5"^^ . _:b437201387 . _:b437200432 "5"^^ . _:b437200282 . _:b437201959 . . _:b437200435 "5"^^ . _:b437200824 . . . _:b437200434 "5"^^ . . _:b437200445 "5"^^ . _:b8124691 . _:b437200814 . _:b437201894 . _:b437200444 "5"^^ . _:b437200857 . . _:b437200447 "5"^^ . _:b437200446 "5"^^ . _:b437201047 . _:b437201835 . _:b437200695 . _:b437200441 "5"^^ . _:b437200780 . _:b437200440 "5"^^ . _:b437201000 . . _:b437200443 "5"^^ . _:b437200442 "5"^^ . _:b437200366 . . _:b437200453 "5"^^ . . _:b437200452 "5"^^ . . _:b437200455 "5"^^ . _:b437200454 "5"^^ . _:b437201277 . _:b437201925 "2"^^ . _:b8124671 "the presence of a large population of human CD4+CCR5+ cells in the intestine, since T cells in the gut-associated lymphoid tissue (GALT) represent a significant population that is targeted by both HIV-1 in humans and SIV in primates (>>28<<\u201334). Overall, the profile of the human cells found in mice receiving ZFN-treated HSC was indistinguishable from mice transplanted with unmodified HSC, both in terms of the percent human cells in each tissue, as well as the frequencies of" . _:b437200449 "5"^^ . . _:b437201924 "2"^^ . . _:b437201734 . _:b437200448 "5"^^ . _:b437201927 "2"^^ . . _:b437200451 "5"^^ . _:b437201926 "2"^^ . _:b437200450 "5"^^ . _:b437201921 "2"^^ . _:b437201869 . . _:b437200461 "5"^^ . _:b437201920 "2"^^ . _:b8124695 . . _:b437201051 . _:b437200460 "5"^^ . . _:b437201923 "2"^^ . . . _:b437200463 "5"^^ . _:b8124717 . _:b437201922 "2"^^ . _:b8124693 . . _:b437200462 "5"^^ . _:b437201933 "2"^^ . _:b437201987 . _:b437200457 "5"^^ . _:b437200983 . . _:b437201932 "2"^^ . . . _:b437200456 "5"^^ . _:b8124721 . . _:b437201935 "2"^^ . _:b437200459 "5"^^ . . _:b437201934 "2"^^ . _:b437200458 "5"^^ . . _:b437201929 "2"^^ . _:b437201083 . . _:b437200542 . _:b437200469 "5"^^ . _:b437200515 . . _:b437201928 "2"^^ . . _:b437201285 . . _:b437200468 "5"^^ . . _:b437201931 "2"^^ . _:b437200471 "5"^^ . . _:b437201930 "2"^^ . . _:b437200470 "5"^^ . . . _:b437201941 "2"^^ . _:b437201881 . _:b437200465 "5"^^ . _:b437201940 "2"^^ . _:b437200634 . _:b437200464 "5"^^ . . _:b437201943 "2"^^ . _:b437200467 "5"^^ . _:b437200439 . _:b437201758 . . _:b437201942 "2"^^ . _:b437200466 "5"^^ . _:b437201937 "2"^^ . _:b437200477 "5"^^ . _:b437201936 "2"^^ . _:b437200476 "5"^^ . _:b437201939 "2"^^ . . _:b437200479 "5"^^ . _:b437201938 "2"^^ . _:b437200478 "5"^^ . _:b437201949 "2"^^ . _:b437200473 "5"^^ . _:b437200781 . _:b437201948 "2"^^ . _:b437200472 "5"^^ . _:b437201951 "2"^^ . . _:b437200475 "5"^^ . . . _:b437201950 "2"^^ . _:b437200474 "5"^^ . _:b437200621 . _:b437201945 "2"^^ . . _:b437200485 "5"^^ . . _:b8124696 . _:b437201944 "2"^^ . _:b437200484 "5"^^ . _:b437201947 "2"^^ . . _:b437200487 "5"^^ . _:b437200826 . _:b437201946 "2"^^ . . _:b437200486 "5"^^ . _:b437201957 "2"^^ . _:b8124685 . _:b8124644 . _:b437200481 "5"^^ . . . _:b437201956 "2"^^ . _:b437200480 "5"^^ . _:b437201959 "2"^^ . _:b437200483 "5"^^ . _:b437200697 . _:b437201958 "2"^^ . _:b437200482 "5"^^ . . . _:b437201953 "2"^^ . _:b437200493 "5"^^ . _:b437201952 "2"^^ . _:b437200793 . _:b437200492 "5"^^ . _:b437201955 "2"^^ . _:b437200876 . _:b437200495 "5"^^ . _:b437201367 . _:b437201954 "2"^^ . _:b437201815 . _:b437200494 "5"^^ . _:b437200756 . _:b437201965 "2"^^ . _:b437200489 "5"^^ . _:b437200712 . _:b437201964 "2"^^ . _:b437200488 "5"^^ . _:b437201967 "2"^^ . _:b437200491 "5"^^ . . _:b437201966 "2"^^ . _:b437200490 "5"^^ . _:b437201333 . _:b437200664 . _:b437201961 "2"^^ . _:b437200501 "5"^^ . _:b437201169 . _:b437200559 . _:b437201960 "2"^^ . _:b437200500 "5"^^ . _:b437200603 . _:b437201963 "2"^^ . _:b437201198 . _:b437200503 "5"^^ . _:b437201962 "2"^^ . _:b437200502 "5"^^ . _:b437201973 "2"^^ . . _:b437200497 "5"^^ . _:b437201134 . _:b437201252 . _:b437201972 "2"^^ . . _:b437201530 . _:b437200496 "5"^^ . _:b437201975 "2"^^ . _:b437200499 "5"^^ . . _:b437201974 "2"^^ . _:b437200691 . _:b437200498 "5"^^ . . _:b437201969 "2"^^ . . _:b437200509 "5"^^ . _:b437201952 . _:b437201968 "2"^^ . . _:b437200508 "5"^^ . _:b437201102 . _:b437201971 "2"^^ . _:b437200511 "5"^^ . _:b437201970 "2"^^ . _:b437200510 "5"^^ . _:b437200354 . _:b437201981 "2"^^ . _:b437200505 "5"^^ . _:b437201920 . _:b437201980 "2"^^ . _:b437200504 "5"^^ . _:b437201983 "2"^^ . . _:b437200507 "5"^^ . _:b437201327 . _:b437201982 "2"^^ . _:b437201971 . _:b437200506 "5"^^ . _:b437201977 "2"^^ . _:b437200517 "5"^^ . _:b437201338 . . _:b437201976 "2"^^ . . . _:b437200516 "5"^^ . _:b437200414 . _:b437201979 "2"^^ . . _:b437201822 . _:b437200519 "5"^^ . _:b437201978 "2"^^ . _:b437200518 "5"^^ . . . _:b437201989 "2"^^ . . _:b437200962 . _:b437200513 "5"^^ . _:b437201988 "2"^^ . _:b437200512 "5"^^ . . _:b437201991 "2"^^ . _:b437200515 "5"^^ . . _:b437201990 "2"^^ . _:b437201595 . _:b437200514 "5"^^ . _:b437201985 "2"^^ . _:b437201546 . _:b437200525 "5"^^ . _:b8124698 . _:b437201984 "2"^^ . _:b437200524 "5"^^ . . _:b437201987 "2"^^ . _:b437200527 "5"^^ . _:b437201654 . _:b437201986 "2"^^ . _:b437200526 "5"^^ . . . _:b437201997 "2"^^ . _:b437200521 "5"^^ . . _:b437201996 "2"^^ . _:b437200520 "5"^^ . _:b437201015 . _:b437200905 . _:b437201999 "2"^^ . _:b8124651 . _:b437200523 "5"^^ . _:b437200582 . _:b437201998 "2"^^ . . _:b437200522 "5"^^ . . _:b437201993 "2"^^ . . _:b437200533 "5"^^ . . _:b437201992 "2"^^ . _:b437200939 . _:b437200532 "5"^^ . _:b437201995 "2"^^ . . _:b437200535 "5"^^ . _:b437201994 "2"^^ . . _:b437200534 "5"^^ . . . _:b437202005 "2"^^ . _:b8124717 "In addition, the experience of autologous HSC transplantation in gene therapy for patients with genetic diseases such as ADA-SCID (59\u201360), chronic granulomatous disease (>>61<<) and X-linked adrenoleukodystrophy (62), is that non-myeloablative conditioning can be used to facilitate engraftment of gene-modified autologous HSC with minimal associated toxicity." . _:b437200529 "5"^^ . _:b437201097 . _:b437202004 "2"^^ . . _:b437200528 "5"^^ . . _:b437202007 "2"^^ . . _:b437200531 "5"^^ . _:b437202006 "2"^^ . _:b8124658 . . _:b437200530 "5"^^ . _:b437202001 "2"^^ . _:b437200203 . _:b437200541 "5"^^ . _:b437201492 . _:b437202000 "2"^^ . _:b437200540 "5"^^ . . _:b437202003 "2"^^ . _:b437200543 "4"^^ . _:b437202002 "2"^^ . _:b8124641 . _:b437200880 . . _:b437200542 "5"^^ . . . _:b437200537 "5"^^ . _:b437201328 . . _:b437200620 . _:b437200536 "5"^^ . . _:b437201910 . _:b437201978 . . _:b437200539 "5"^^ . _:b437201596 . _:b437200538 "5"^^ . . _:b437201790 . _:b437200549 "4"^^ . _:b437202008 "2"^^ . _:b437200548 "4"^^ . . _:b437200551 "4"^^ . _:b437201135 . . _:b437201923 . . _:b437200550 "4"^^ . . _:b437201072 . _:b437200545 "4"^^ . . . _:b437201232 . _:b437200544 "4"^^ . . . _:b437200547 "4"^^ . _:b437200546 "4"^^ . _:b437200557 "4"^^ . _:b437200262 . _:b437200556 "4"^^ . . _:b8124725 . _:b437200559 "4"^^ . . . . _:b437200558 "4"^^ . . _:b437201076 . _:b437200349 . . _:b437200553 "4"^^ . . . . _:b437200552 "4"^^ . . _:b437200555 "4"^^ . _:b437200975 . _:b437200554 "4"^^ . . . . _:b437200565 "4"^^ . _:b437200564 "4"^^ . _:b437200854 . . _:b437200567 "4"^^ . . . _:b437200670 . _:b437200566 "4"^^ . _:b437201068 . . _:b437200561 "4"^^ . _:b437201605 . _:b437201787 . _:b437200574 . _:b437201882 . . _:b437200560 "4"^^ . . . _:b437201035 . . _:b437200563 "4"^^ . _:b437200872 . _:b437201964 . . . _:b437200562 "4"^^ . . _:b437200573 "4"^^ . . . _:b437200572 "4"^^ . _:b437201304 . . _:b437200575 "4"^^ . . . _:b437200351 . _:b437200708 . _:b437200574 "4"^^ . . _:b437200569 "4"^^ . . _:b437200568 "4"^^ . _:b437200822 . _:b437200571 "4"^^ . _:b437200896 . _:b437200570 "4"^^ . . _:b437200581 "4"^^ . _:b437201455 . . _:b437201756 . . . _:b437200580 "4"^^ . _:b437200902 . _:b437201800 . _:b437200583 "4"^^ . _:b437200582 "4"^^ . _:b437200345 . _:b437201246 . . _:b437200577 "4"^^ . _:b437200576 "4"^^ . _:b437200579 "4"^^ . . _:b437201634 . . _:b437200863 . _:b437201403 . . . _:b437201046 . _:b437200578 "4"^^ . _:b437200589 "4"^^ . . _:b437201620 . _:b437200588 "4"^^ . . _:b437200591 "4"^^ . . _:b437200590 "4"^^ . . _:b437200585 "4"^^ . _:b437200682 . _:b437200584 "4"^^ . _:b437200759 . . _:b437201907 . _:b437200587 "4"^^ . _:b437201400 . _:b437200283 . . _:b437200825 . _:b437200586 "4"^^ . _:b437200597 "4"^^ . . . _:b437201060 . _:b437200596 "4"^^ . _:b437200527 . _:b437200599 "4"^^ . _:b437201583 . _:b437200598 "4"^^ . _:b437200459 . _:b437201741 . . _:b437200593 "4"^^ . _:b437200592 "4"^^ . _:b437200257 . _:b437201002 . _:b437200724 . _:b437200595 "4"^^ . . _:b437200629 . _:b437200594 "4"^^ . . _:b437200605 "4"^^ . _:b437200604 "4"^^ . . . . _:b437200607 "4"^^ . . . . _:b437200606 "4"^^ . _:b437200601 "4"^^ . _:b437200600 "4"^^ . _:b437200344 . _:b437200603 "4"^^ . _:b437200353 . . . _:b437200602 "4"^^ . . _:b437201903 . . _:b437200613 "4"^^ . _:b437200612 "4"^^ . _:b437200668 . _:b437201027 . _:b437200615 "4"^^ . _:b437201438 . _:b437200614 "4"^^ . _:b437201821 . _:b437200321 . _:b437200609 "4"^^ . . _:b437201700 . _:b437200608 "4"^^ . _:b437200611 "4"^^ . . _:b437201807 . . _:b437200610 "4"^^ . _:b437200621 "4"^^ . . _:b437200620 "4"^^ . _:b437200623 "4"^^ . _:b437200202 . . . _:b437200789 . . _:b437200622 "4"^^ . _:b437200617 "4"^^ . _:b437200663 . _:b437200616 "4"^^ . _:b437200619 "4"^^ . _:b437200525 . _:b437200847 . _:b437200618 "4"^^ . . . _:b437200629 "4"^^ . _:b437200846 . . _:b437200328 . _:b437200628 "4"^^ . _:b437200631 "4"^^ . _:b437200829 . _:b437200630 "4"^^ . _:b437200625 "4"^^ . . _:b437200624 "4"^^ . _:b437200410 . . _:b437200757 . _:b437200627 "4"^^ . _:b437201082 . _:b437200626 "4"^^ . . _:b437200637 "4"^^ . . _:b437200636 "4"^^ . _:b437200639 "4"^^ . _:b437201184 . . _:b437200638 "4"^^ . _:b437200296 . _:b437200633 "4"^^ . . _:b437201140 . _:b437200632 "4"^^ . . _:b437200469 . _:b437200635 "4"^^ . . . _:b437200634 "4"^^ . _:b437200645 "4"^^ . . _:b437200644 "4"^^ . . _:b437200647 "4"^^ . _:b437200646 "4"^^ . _:b437201557 . . . _:b437201303 . _:b437200641 "4"^^ . _:b437200640 "4"^^ . . _:b437200643 "4"^^ . _:b437200961 . . . _:b437200642 "4"^^ . . _:b437200653 "4"^^ . _:b437201183 . . _:b437200652 "4"^^ . . . _:b437200655 "4"^^ . _:b437200654 "4"^^ . _:b437200649 "4"^^ . _:b437200988 . _:b437200648 "4"^^ . . _:b437200651 "4"^^ . _:b437200650 "4"^^ . _:b437200293 . _:b437200661 "4"^^ . . . . _:b437200660 "4"^^ . _:b437200536 . . _:b437200663 "4"^^ . _:b437201203 . _:b437200662 "4"^^ . _:b437201548 . . . _:b437201368 . . _:b437200657 "4"^^ . . . _:b437200656 "4"^^ . . . _:b437200659 "4"^^ . . _:b437200658 "4"^^ . _:b437200765 . _:b437200669 "4"^^ . . . _:b437200668 "4"^^ . _:b437201188 . . _:b437200671 "4"^^ . _:b437201470 . . _:b437200670 "4"^^ . _:b437200665 "4"^^ . _:b437200664 "4"^^ . _:b437201783 . _:b437200667 "4"^^ . . . . . _:b437201541 . _:b437200666 "4"^^ . _:b437200677 "4"^^ . . _:b437200676 "4"^^ . _:b437201491 . _:b437200679 "4"^^ . . . _:b437200678 "4"^^ . _:b437200673 "4"^^ . . _:b437201205 . _:b437201486 . _:b8124645 "with a lack of CCR5 (5\u20137) has spurred the development of therapies aimed at blocking the virus-CCR5 interaction, and CCR5 antagonists have proved to be an effective salvage therapy in patients with drug-resistant strains of HIV-1 (>>8<<)." . . _:b437200672 "4"^^ . _:b437200700 . . _:b437200675 "4"^^ . . . _:b437200674 "4"^^ . _:b437201694 . . _:b437201713 . _:b437201062 . . _:b437201633 . _:b437200685 "4"^^ . . _:b437201248 . _:b437200684 "4"^^ . _:b437200221 . _:b437200687 "4"^^ . . _:b437200686 "4"^^ . _:b8124711 . _:b437200681 "4"^^ . _:b437200680 "4"^^ . _:b437201685 . _:b437200683 "4"^^ . . _:b437200682 "4"^^ . _:b8124672 "This strain of HIV-1 has previously been used in humanized mouse models, where it causes a robust infection and significant CD4+ T cell depletion (>>35<<,36). These observations mimic the human infection, where depletion of CD4+ CCR5+ lymphocytes results from a combination of direct infection, systemic immune activation and the upregulation of CCR5 on thymocytes (36,37,38). Following" . _:b437200693 "4"^^ . _:b437200692 "4"^^ . _:b437200695 "4"^^ . . . _:b437200694 "4"^^ . _:b437200362 . . _:b437201771 . _:b437200689 "4"^^ . . _:b437200688 "4"^^ . _:b437200691 "4"^^ . _:b8124657 . _:b8124685 "methods" . _:b437200690 "4"^^ . _:b437200701 "4"^^ . _:b437200700 "4"^^ . _:b437201363 . _:b437200358 . _:b437200703 "4"^^ . . . _:b437200702 "4"^^ . . . . _:b437200697 "4"^^ . . . _:b437200696 "4"^^ . _:b8124660 . . _:b437200699 "4"^^ . . _:b437200698 "4"^^ . _:b437200709 "4"^^ . _:b437201281 . . _:b437200708 "4"^^ . _:b437201294 . _:b437200711 "4"^^ . _:b8124667 . . . _:b437200710 "4"^^ . _:b8124666 . _:b437200705 "4"^^ . . . _:b437200704 "4"^^ . . _:b437200707 "4"^^ . . _:b8124675 . _:b437200706 "4"^^ . _:b437200717 "4"^^ . _:b437201070 . _:b437200716 "4"^^ . . _:b437200719 "4"^^ . . _:b437200718 "4"^^ . . _:b437200713 "4"^^ . . . _:b437200712 "4"^^ . _:b8124648 . _:b437200715 "4"^^ . _:b437200714 "4"^^ . _:b437201254 . _:b437201958 . _:b8124650 . . _:b8124663 . . _:b437200725 "4"^^ . _:b437200724 "4"^^ . _:b437200802 . _:b437200727 "3"^^ . _:b437201905 . _:b437200726 "3"^^ . _:b437200721 "4"^^ . _:b437200720 "4"^^ . . _:b8124696 _:b8124700 . _:b437200723 "4"^^ . _:b8124696 _:b8124701 . _:b8124696 _:b8124702 . _:b8124655 . _:b8124696 _:b8124703 . _:b437200722 "4"^^ . _:b8124696 _:b8124697 . _:b8124696 _:b8124698 . . _:b8124696 _:b8124699 . _:b437200733 "3"^^ . _:b8124696 _:b8124708 . _:b8124696 _:b8124709 . _:b8124723 "Interestingly, the patient receiving the CCR5\u039432 HSC transplantation harbored CXCR4-utilizing viruses before the procedure, yet he did not experience a tropism switch, and instead, successfully controlled his HIV-1 infection long-term (>>9<<,10). For the present, similar to the recommendations for the use of CCR5 inhibitor drugs, it may be prudent to restrict CCR5 ZFN treatment of HSC to individuals with no detectable CXCR4-tropic viruses." . . _:b437201991 . _:b8124696 _:b8124710 . _:b8124696 _:b8124711 . _:b437200732 "3"^^ . _:b8124696 _:b8124704 . _:b8124696 _:b8124705 . _:b8124696 _:b8124706 . _:b8124696 _:b8124707 . _:b437200735 "3"^^ . _:b437200934 . _:b8124696 _:b8124716 . _:b437200359 . _:b8124696 _:b8124717 . _:b8124696 _:b8124718 . _:b437200537 . _:b8124696 _:b8124719 . _:b437200734 "3"^^ . _:b8124696 _:b8124712 . _:b8124696 _:b8124713 . . _:b8124696 _:b8124714 . _:b8124696 _:b8124715 . _:b437200729 "3"^^ . _:b8124696 _:b8124724 . _:b8124696 _:b8124725 . _:b437200350 . _:b8124696 _:b8124726 . _:b8124696 _:b8124727 . _:b437200728 "3"^^ . _:b8124696 _:b8124720 . _:b8124696 _:b8124721 . _:b8124696 _:b8124722 . _:b8124696 _:b8124723 . _:b437200731 "3"^^ . . _:b437201469 . . _:b8124696 _:b8124728 . _:b437200730 "3"^^ . _:b8124696 _:b8124729 . . . _:b437200450 . _:b437200741 "3"^^ . _:b8124685 _:b8124688 . _:b8124685 _:b8124689 . _:b437200447 . _:b8124685 _:b8124690 . . _:b8124685 _:b8124691 . _:b437200740 "3"^^ . _:b8124685 _:b8124692 . _:b8124685 _:b8124693 . _:b8124685 _:b8124694 . _:b8124685 _:b8124695 . _:b437200743 "3"^^ . . _:b437201885 . _:b437200742 "3"^^ . _:b437200737 "3"^^ . _:b437200736 "3"^^ . _:b437200595 . . _:b437200739 "3"^^ . _:b437201845 . _:b437201805 . _:b437200738 "3"^^ . _:b8124685 _:b8124686 . _:b8124685 _:b8124687 . _:b437200749 "3"^^ . _:b437201767 . _:b437200748 "3"^^ . _:b437200751 "3"^^ . . . _:b437200605 . _:b437200750 "3"^^ . . . _:b437200930 . _:b437200745 "3"^^ . "10.1038%2Fnbt.1663" . _:b437201077 . _:b437200744 "3"^^ . _:b437201092 . _:b437200807 . _:b437200747 "3"^^ . . _:b437200746 "3"^^ . . _:b437201609 . _:b437200757 "3"^^ . . _:b437201312 . _:b437200756 "3"^^ . _:b437201237 . _:b437200759 "3"^^ . . _:b437201168 . _:b437200758 "3"^^ . . . _:b437200543 . . _:b437200753 "3"^^ . . _:b437200752 "3"^^ . _:b437201804 . _:b437200755 "3"^^ . . _:b8124710 "where normal hematopoietic cells have a selective advantage, so that spontaneous monoclonal reversions that have occurred in a small number of patients can lead to the selective outgrowth of these cells and amelioration of symptoms (>>50<<\u201353)." . _:b437200754 "3"^^ . . _:b437200765 "3"^^ . _:b437201626 . _:b437200764 "3"^^ . _:b8124713 . _:b437201550 . _:b437200767 "3"^^ . . _:b437200766 "3"^^ . _:b437200207 . . . _:b437200761 "3"^^ . . _:b437200760 "3"^^ . . _:b437201229 . _:b437200763 "3"^^ . _:b437201044 . . . _:b437200762 "3"^^ . . _:b437201659 . _:b437201034 . _:b437200773 "3"^^ . _:b437201133 . _:b437201572 . . _:b437200772 "3"^^ . . _:b437201555 . _:b437200775 "3"^^ . . _:b437200774 "3"^^ . _:b437200337 . _:b437200769 "3"^^ . . _:b437200768 "3"^^ . . . _:b437200427 . . _:b437200771 "3"^^ . _:b437200770 "3"^^ . . _:b437200505 . . _:b437200781 "3"^^ . . _:b437201672 . _:b437200780 "3"^^ . _:b437200276 . _:b437200783 "3"^^ . _:b437200614 . _:b437201048 . _:b437200782 "3"^^ . _:b437200777 "3"^^ . _:b437201271 . _:b437200776 "3"^^ . _:b437200779 "3"^^ . _:b437200229 . _:b437200778 "3"^^ . _:b437200789 "3"^^ . _:b437201781 . _:b437201462 . _:b437200197 . _:b437200788 "3"^^ . _:b437200438 . _:b437201108 . . _:b437200347 . _:b437200791 "3"^^ . _:b437200790 "3"^^ . _:b437201990 . _:b437200785 "3"^^ . . . _:b437200784 "3"^^ . . . _:b437200706 . _:b437200787 "3"^^ . _:b437200786 "3"^^ . . _:b437200797 "3"^^ . _:b437200796 "3"^^ . . _:b8124724 . . _:b437200799 "3"^^ . _:b437200916 . _:b437200798 "3"^^ . _:b437200793 "3"^^ . . _:b437200792 "3"^^ . _:b437200612 . _:b437200795 "3"^^ .