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- Title
Highly efficient endogenous human gene correction using designed zinc-finger nucleases.
- Authors
Urnov, Fyodor D; Miller, Jeffrey C; Lee, Ya-Li; Beausejour, Christian M; Rock, Jeremy M; Augustus, Sheldon; Jamieson, Andrew C; Porteus, Matthew H; Gregory, Philip D; Holmes, Michael C
- 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 IL2Rgamma 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.
- Publication
Nature, 2005, Vol 435, Issue 7042, p646
- ISSN
1476-4687
- Publication type
Journal Article
- DOI
10.1038/nature03556