BRCA1 deficient embryonic stem cells display a decreased homologous recombination frequency and an increased frequency of non-homologous recombination that is corrected by expression of a Brca1 transgene
| Autor: | Andrew Xiao, Amy R. Mohn, John N. Snouwaert, Anne M. Latour, Beverly H. Koller, Lauren M. DiBiase, Lori C. Gowen |
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| Rok vydání: | 1999 |
| Předmět: |
Cancer Research
endocrine system diseases DNA repair Transgene Restriction Mapping Genes BRCA1 RAD51 Breast Neoplasms Mice Transgenic Biology Mice Risk Factors Genetics Animals Humans skin and connective tissue diseases Molecular Biology Gene Cells Cultured Mice Knockout Ovarian Neoplasms Recombination Genetic Reverse Transcriptase Polymerase Chain Reaction Stem Cells Wild type Gene targeting Embryo Mammalian Molecular biology Female Stem cell Homologous recombination Gene Deletion |
| Zdroj: | Oncogene. 18:7900-7907 |
| ISSN: | 1476-5594 0950-9232 |
| Popis: | BRCA1 is a nuclear phosphoprotein that has been classified as a tumor suppressor based on the fact that women carrying a mutated copy of the BRCA1 gene are at increased risk of developing breast and ovarian cancer. The association of BRCA1 with RAD51 has led to the hypothesis that BRCA1 is involved in DNA repair. We describe here the generation and analysis of murine embryonic stem (ES) cell lines in which both copies of the murine homologue of the human BRCA1 gene have been disrupted by gene targeting. We show that exogenous DNA introduced into these BRCA1 deficient cells by electroporation is randomly integrated into the genome at a significantly higher rate than in wild type ES cells. In contrast, integration of exogenous DNA by homologous recombination occurs in BRCA1 deficient cells at a significantly lower rate than in wild type controls. When BRCA1 expression is re-established at 5-10% of normal levels by introduction of a Brca1 transgene into BRCA1 deficient ES cells, the frequency of random integration is reduced to wild type levels, although the frequency of homologous recombination is not significantly improved. These results suggest that BRCA1 plays a role in determining the response of cells to double stranded DNA breaks. |
| Databáze: | OpenAIRE |
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