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An improved zinc-finger nuclease architecture for highly specific genome editing
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An improved zinc-finger nuclease architecture for highly specific genome editing
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Journal Article
An improved zinc-finger nuclease architecture for highly specific genome editing
2007
Overview
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.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering/Biotechnology
/ Deoxyribonucleases, Type II Site-Specific - chemistry
/ DNA
/ Fundamental and applied biological sciences. Psychology
/ Genome
/ Genomics
/ Green Fluorescent Proteins - chemistry
/ Humans
/ Methods. Procedures. Technologies
/ Reagents
/ Zinc
