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Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1
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Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1

Article DOI: 10.3791/2468-v • 08:53 min • February 17th, 2011
February 17th, 2011 •
1, 2, 1
1Department of Biomedical Informatics, The Ohio State University, 2Departments of Molecular Immunology and Clinical Oncology, Tohoku University

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Summary
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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.

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BRCA1 Mutations Homologous Recombination Endogenous BRCA1 Missense Mutations Functional Analysis Cell Lines RNAi Targeting BRCA1 MRNA BRCA1 Variant Acute Silencing Secondary Mutations Compensation For Loss Of Function HeLa-derived Cell Line Homologous Recombination Assay Recombination Substrate I-SceI Restriction Enzyme Site GFP Allele
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