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Published byMelissa Allison Modified over 5 years ago
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BRCA1 sequence variants classified as pathogenic do not restore HR
BRCA1 sequence variants classified as pathogenic do not restore HR. R26CreERT2/hBRCA1;Brca1SCo/Δ;Pim1DR-GFP/wt embryonic stem cells carrying the DR-GFP reporter gene in the Pim1 locus and mutant BRCA1 or controls in the Rosa26 locus were switched using 4-OH... BRCA1 sequence variants classified as pathogenic do not restore HR. R26CreERT2/hBRCA1;Brca1SCo/Δ;Pim1DR-GFP/wt embryonic stem cells carrying the DR-GFP reporter gene in the Pim1 locus and mutant BRCA1 or controls in the Rosa26 locus were switched using 4-OHT and transfected with a vector expressing I-SceI and mCherry. Transfected cells were analyzed for GFP expression as a measure of HR activity. Expression of BRCA1 wild-type (WT) cDNA resulted in increased HR compared to the empty RMCE vector (Vector) control. Error bars indicate the SD between the results of three independent I-SceI transfections. Significantly decreased HR activity compared with the wild-type control (green line) is indicated. Peter Bouwman et al. Cancer Discovery 2013;3: ©2013 by American Association for Cancer Research
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