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Curr Protoc Mol Biol. 2011 Oct;Chapter 12:Unit12.13. doi: 10.1002/0471142727.mb1213s96.

Engineering designer nucleases with customized cleavage specificities.

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1
Molecular Pathology Unit, Center for Cancer Research, and Center for Computational and Integrative Biology, Massachusetts General Hospital, Charlestown, Massachusetts, USA.

Abstract

Engineered designer nucleases can be used to efficiently modify genomic sequence in a wide variety of model organisms and cell types. Zinc finger nucleases (ZFNs), consisting of an engineered zinc finger array fused to a non-specific cleavage domain, have been extensively used to modify a broad range of endogenous genes. Protocols for engineering ZFNs targeted to specific gene sequences of interest using the context-dependent assembly (CoDA) method are described in this unit.

PMID:
21987054
PMCID:
PMC3379964
DOI:
10.1002/0471142727.mb1213s96
[Indexed for MEDLINE]
Free PMC Article
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