An improved zinc-finger nuclease architecture for highly specific genome editing View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2007-07

AUTHORS

Jeffrey C Miller, Michael C Holmes, Jianbin Wang, Dmitry Y Guschin, Ya-Li Lee, Igor Rupniewski, Christian M Beausejour, Adam J Waite, Nathaniel S Wang, Kenneth A Kim, Philip D Gregory, Carl O Pabo, Edward J Rebar

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. More... »

PAGES

778-785

Journal

TITLE

Nature Biotechnology

ISSUE

7

VOLUME

25

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt1319

    DOI

    http://dx.doi.org/10.1038/nbt1319

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1002154526

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/17603475


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