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An improved zinc-finger nuclease architecture for highly specific genome editing.
- Source :
- Nature Biotechnology; Jul2007, Vol. 25 Issue 7, p778-785, 8p, 5 Diagrams, 1 Graph
- Publication Year :
- 2007
-
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. [ABSTRACT FROM AUTHOR]
- Subjects :
- NUCLEASES
GENES
DNA
DIMERS
NUCLEIC acids
Subjects
Details
- Language :
- English
- ISSN :
- 10870156
- Volume :
- 25
- Issue :
- 7
- Database :
- Complementary Index
- Journal :
- Nature Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 25681874
- Full Text :
- https://doi.org/10.1038/nbt1319