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- W4200540919 abstract "DNA double-strand breaks are the most dangerous DNA lesions that may lead to massive loss of genetic information and cell death. Cells repair DSBs using two major pathways: nonhomologous end joining (NHEJ) and homologous recombination (HR). Perturbations of NHEJ and HR are often associated with premature aging and tumorigenesis, hence it is important to have a quantitative way of measuring each DSB repair pathway. Our laboratory has developed fluorescent reporter constructs that allow sensitive and quantitative measurement of NHEJ and HR. The constructs are based on an engineered GFP gene containing recognition sites for a rare-cutting I-SceI endonuclease for induction of DSBs. The starting constructs are GFP negative as the GFP gene is inactivated by an additional exon, or by mutations. Successful repair of the I-SceI-induced breaks by NHEJ or HR restores the functional GFP gene. The number of GFP positive cells counted by flow cytometry provides quantitative measure of NHEJ or HR efficiency." @default.
- W4200540919 created "2021-12-31" @default.
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- W4200540919 creator A5052978985 @default.
- W4200540919 date "2010-09-08" @default.
- W4200540919 modified "2023-09-25" @default.
- W4200540919 title "Analysis of DNA Double-strand Break (DSB) Repair in Mammalian Cells" @default.
- W4200540919 doi "https://doi.org/10.3791/2002-v" @default.
- W4200540919 hasPublicationYear "2010" @default.
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