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

Autor: Yuan J; Merck Research Laboratories, Boston, MA 02115, USA., Strack PR, Toniatti C, Pelletier M
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2013 Mar 01; Vol. 434 (1), pp. 96-8. Date of Electronic Publication: 2012 Nov 10.
DOI: 10.1016/j.ab.2012.11.002
Abstrakt: 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.
(Copyright © 2012 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: