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Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery.
- Source :
-
Nature Biotechnology . Nov2007, Vol. 25 Issue 11, p1298-1306. 9p. 2 Diagrams, 4 Graphs. - Publication Year :
- 2007
-
Abstract
- Achieving the full potential of zinc-finger nucleases (ZFNs) for genome engineering in human cells requires their efficient delivery to the relevant cell types. Here we exploited the infectivity of integrase-defective lentiviral vectors (IDLV) to express ZFNs and provide the template DNA for gene correction in different cell types. IDLV-mediated delivery supported high rates (13–39%) of editing at the IL-2 receptor common γ-chain gene (IL2RG) across different cell types. IDLVs also mediated site-specific gene addition by a process that required ZFN cleavage and homologous template DNA, thus establishing a platform that can target the insertion of transgenes into a predetermined genomic site. Using IDLV delivery and ZFNs targeting distinct loci, we observed high levels of gene addition (up to 50%) in a panel of human cell lines, as well as human embryonic stem cells (5%), allowing rapid, selection-free isolation of clonogenic cells with the desired genetic modification. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 10870156
- Volume :
- 25
- Issue :
- 11
- Database :
- Academic Search Index
- Journal :
- Nature Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 27404303
- Full Text :
- https://doi.org/10.1038/nbt1353