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- Title
Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery.
- Authors
Lombardo, Angelo; Genovese, Pietro; Beausejour, Christian M.; Colleoni, Silvia; Ya-Li Lee; Kim, Kenneth A.; Ando, Dale; Urnov, Fyodor D.; Galli, Cesare; Gregory, Philip D.; Holmes, Michael C.; Naldini, Luigi
- 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.
- Subjects
GENETICS; GENES; EMBRYONIC stem cells; STEM cells; GENOMES; CELLS; DNA; NUCLEIC acids; TRANSGENES
- Publication
Nature Biotechnology, 2007, Vol 25, Issue 11, p1298
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt1353