HCT 116 BRCA2 -/- 46 Cell Line
HCT116 BRCA2 -/- [46] human colorectal carcinoma cell line generated to study the role of BRCA2 in DNA repair and a range of cancers.
Contributors
| Inventor | Institute |
|---|---|
| Carlos Caldas | Cancer Research UK Cambridge Institute |
| Cat. #: | 153531 |
|---|---|
| Tool sub type: | Continuous |
| Unit size: | 1×10^6 cells / vial |
| Cancer types: | Digestive / Gastrointestinal cancer |
| Research Fields: | Cancer;Drug development;Genetics |
| Organism: | Human |
| Tissue: | Colon |
| Model: | Knock-Out |
| Morphology: | Polygonal epithelium |
| Growth properties: | Adherent |
| Cancer Types In Detail: | Colorectal carcinoma |
| Primary citation: | Xu et al. 2014. J Pathol. 234(3):386-97. PMID: 25043256 |
| Alternate name: | HCT 116 BRCA2(-/-) clone 46; HCT 116 BRCA2(-/-) |
|---|---|
| Product description: | BRAC2 inherited mutations predispose carriers to various early onset cancers, including breast and ovarian. HCT116 BRCA2 -/- human colorectal carcinoma cell line was generated to study the role of BRCA2 in DNA repair, investigate the effects of BRCA2 mutations in a range of cancers, and identify additional functions of the BRCA2 gene. This includes the upregulation and effect of interferon-related genes (including APOBEC3F and APOBEC3G) when the BRAC2 gene is lost. This homozygous knockout cell line shows phenotypes consistent with previous reports, including loss of Rad51 foci in the presence of double-strand breaks, chromosomal rearrangements and elevated sensitivity to the DNA-damaging agents phleomycin and Parp1 inhibitors. This cell line has been used to show DNA damage occurs in BRCA2 knockout HCT116 cells even in the absence of exogenous genotoxic stress |
| CRISPR: | No |
| Conditional: | No |
| Production details: | Targeted disruption of the human BRCA2 locus in HCT116 cells by homologous recombination. The gene targeting construct was generated by using a recombinant adeno-associated virus (rAAV) system. The deletion was confirmed by Southern Blot and Western Blot using antibodies against the C-terminus of the BRCA2 protein. This cell line was noted as clone 46 by the originating laboratory |
| Additional notes: | Clone 42 was generated at the same time as clone 46 cited in Xu et al. 2014. J Pathol. 234(3):386-97. PMID: 25043256. The originating laboratory have advised they observed no difference in culture phenotype between clone 42 and clone 46. |
| Cellosaurus ID: | CVCL_VP26 |
| Parental cell line: | HCT 116 cell line |
| Model description: | Targeted disruption of human BRCA2 by homologous recombination |
| disease: | Cancer |
| Format: | Frozen |
|---|---|
| Storage conditions: | Liquid Nitrogen |
| Shipping conditions: | Dry ice |
| Growth medium: | McCoy's 5a medium modified, with 10% foetal bovine serum (FBS) and 2mM L-Glutamine |
| Subculture routine: | Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 2-4×10⁴ cells/cm² using 0.05% trypsin/EDTA solution. Cells are cultured at 37°C, in 95% air and 5% CO₂, and passaged every 3 to 4 days. |
| Recommended controls: | HCT 116 parental line |
| Mycoplasma free: | Yes |
| Biosafety level: | 1 |
| References: |
Xu et al. 2014. J Pathol. 234(3):386-97. PMID: 25043256 Issaeva et al. 2010. Cancer Res. 70(15):6268-76. PMID: 20631063 |
|---|
Images
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Product Images
HCT 116 BRCA2 -/- 46 Cell Line. Early log phase.
HCT 116 BRCA2 -/- 46 Cell Line. Early log phase. Image courtesy of the European Collection of Authenticated Cell Cultures (ECACC), UK.
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