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. 2013 Mar 1;434(1):96-8.
doi: 10.1016/j.ab.2012.11.002. Epub 2012 Nov 10.

A zinc finger nuclease assay to rapidly quantitate homologous recombination proficiency in human cell lines

Jing Yuan  1 , Peter R Strack, Carlo Toniatti, Marc Pelletier
Affiliations

A zinc finger nuclease assay to rapidly quantitate homologous recombination proficiency in human cell lines

Jing Yuan et al. Anal Biochem. .

Abstract

Homologous recombination (HR) is a cellular mechanism for accurate repair of double-strand DNA breaks, often deregulated in cancer. Development of novel cancer therapeutics targeting HR pathways would benefit from a quantitative and rapid means of measuring HR. Here, we describe a zinc finger nuclease (ZFN) assay that can quantify HR. Knockdown of BRCA1 or inactivation of BRCA2 decreased HR activity in cells, whereas gene restoration induced activity 8-fold. HR activity was also reflective of BRCA1/2 status in cells with known endogenous mutations.

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