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

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 nuclea...

Ausführliche Beschreibung

Gespeichert in:

Bibliographische Detailangaben
Veröffentlicht in:	Analytical biochemistry 2013-03, Vol.434 (1), p.96-98
Hauptverfasser:	Yuan, Jing, Strack, Peter R., Toniatti, Carlo, Pelletier, Marc
Format:	Artikel
Sprache:	eng
Schlagworte:	BRCA1 Protein - antagonists & inhibitors BRCA1 Protein - genetics BRCA1 Protein - metabolism BRCA2 Protein - genetics BRCA2 Protein - metabolism Cell Line, Tumor cells Colony formation assay DNA damage Endonucleases - metabolism Enzyme Assays genes HeLa Cells Homologous Recombination human cell lines Humans mutation PCR recombination RNA Interference RNA, Small Interfering - metabolism therapeutics tumor suppressor proteins zinc finger motif Zinc finger nuclease Zinc Fingers
Online-Zugang:	Volltext
Tags:	Tag hinzufügen Keine Tags, Fügen Sie den ersten Tag hinzu!

Beschreibung
Zusammenfassung:	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.
ISSN:	0003-2697 1096-0309
DOI:	10.1016/j.ab.2012.11.002