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| Highly efficient endogenous human gene correction using designed zinc-finger nucleases |
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| Author(s): Urnov FD, Miller JC, Lee YL, Beausejour CM, Rock JM, Augustus S, Jamieson AC, Porteus MH, Gregory PD, Holmes MC |
| Source: NATURE Volume: 435 Issue: 7042 Pages: 646-651 Published: JUN 2 2005 |
| Times Cited: 219 References: 39 |
| Abstract: Permanent modification of the human genome in vivo is impractical owing to the low frequency of homologous recombination in human cells, a fact that hampers biomedical research and progress towards safe and effective gene therapy. Here we report a general solution using two fundamental biological processes: DNA recognition by C2H2 zinc-finger proteins and homology-directed repair of DNA double-strand breaks. Zinc-finger proteins engineered to recognize a unique chromosomal site can be fused to a nuclease domain, and a double-strand break induced by the resulting zinc-finger nuclease can create specific sequence alterations by stimulating homologous recombination between the chromosome and an extrachromosomal DNA donor. We show that zinc-finger nucleases designed against an X-linked severe combined immune deficiency ( SCID) mutation in the IL2R gamma gene yielded more than 18% gene-modified human cells without selection. Remarkably, about 7% of the cells acquired the desired genetic modification on both X chromosomes, with cell genotype accurately reflected at the messenger RNA and protein levels. We observe comparably high frequencies in human T cells, raising the possibility of strategies based on zinc-finger nucleases for the treatment of disease. |
| Document Type: Article |
| Language: English |
| Reprint Address: Holmes, MC (reprint author), Sangamo BioSci Inc, Pt Richmond Tech Ctr 501,Canal Blvd,Suite A100, Richmond, CA 94804 USA |
Addresses:
1. Sangamo BioSci Inc, Richmond, CA 94804 USA
2. Univ Texas, SW Med Ctr, Dept Pediat, Dallas, TX 75390 USA
3. Univ Texas, SW Med Ctr, Dept Biochem, Dallas, TX 75390 USA |
| Publisher: NATURE PUBLISHING GROUP, MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND |
| Subject Category: Multidisciplinary Sciences |
| IDS Number: 931HK |
| ISSN: 0028-0836 |
| DOI: 10.1038/nature03556 |
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