Preparation of Cell Line with an Integrated Recombination Substrate
3:04
Transfection to Deplete Endogenous BRCA1 and add back Mutant BRCA1
5:19
Analysis of Transfectants
6:07
Analysis of the Effects of BRCA1 Mutations on Homologous Recombination via GFP-Expression
7:54
Conclusion
We provide a method for testing BRCA1 variants in a tissue culture based assay for homologous recombination repair of DNA damage by depleting endogenous BRCA1 protein from a cell using RNAi and replacing it with a BRCA1 point mutant that contains a coding change.