title: A zinc finger nuclease assay to rapidly quantitate homologous recombination proficiency in human cell lines.
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.
authors.affiliation: Merck Research Laboratories, Boston, MA 02115, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.forename: Peter R
authors.initials: PR
authors.lastname: Strack
authors.forename: Carlo
authors.initials: C
authors.lastname: Toniatti
authors.forename: Marc
authors.initials: M
authors.lastname: Pelletier
mesh_terms: D019313:BRCA1 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D024682:BRCA2 Protein / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D004720:Endonucleases / Q000378:metabolism*; D057075:Enzyme Assays; D006367:HeLa Cells; D059765:Homologous Recombination; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D019313:BRCA1 Protein; D024682:BRCA2 Protein; D034741:RNA, Small Interfering; D004720:Endonucleases