STUDIES OF BRCA1 PROTEIN USING CRISPR/CAS9 GENE EDITING IN HCC1937 BREAST CANCER CELLS

Abstract

The mutation in the BRCA1 gene in HCC1937 breast cancer cell lines produces a truncated protein that prevents interaction between BRCA1 protein and its partner called CtIP protein. The DNA repair processing needs this interaction to initiate double-strand repair. CRISPR/Cas9 system gene editing will be used to correct this mutation by deleting the insertion of cytosine at position 5382 in the wild-type BRCA1 protein. Co-immunoprecipitation (Co-IP) and western-blot assays will be used to confirm the protein interaction after correction by CRISPR/Cas9 system gene editing. Moreover, DNA repair and IR sensitivity assays will be conducted to study the BRCA1 response in these cell lines. The rate of proliferation and invasion between mutated, corrected and normal cell lines will be measured by using MTT (3-(4,5- dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide) and Transwell assays.