An improved zinc-finger nuclease architecture for highly specific genome editing
- PMID: 17603475
- DOI: 10.1038/nbt1319
An improved zinc-finger nuclease architecture for highly specific genome editing
Abstract
Genome editing driven by zinc-finger nucleases (ZFNs) yields high gene-modification efficiencies (>10%) by introducing a recombinogenic double-strand break into the _targeted gene. The cleavage event is induced using two custom-designed ZFNs that heterodimerize upon binding DNA to form a catalytically active nuclease complex. Using the current ZFN architecture, however, cleavage-competent homodimers may also form that can limit safety or efficacy via off-_target cleavage. Here we develop an improved ZFN architecture that eliminates this problem. Using structure-based design, we engineer two variant ZFNs that efficiently cleave DNA only when paired as a heterodimer. These ZFNs modify a native endogenous locus as efficiently as the parental architecture, but with a >40-fold reduction in homodimer function and much lower levels of genome-wide cleavage. This architecture provides a general means for improving the specificity of ZFNs as gene modification reagents.
Comment in
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Precision genome surgery.Nat Biotechnol. 2007 Jul;25(7):743-4. doi: 10.1038/nbt0707-743. Nat Biotechnol. 2007. PMID: 17621297 No abstract available.
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