CELL LINES
Contributor Information
- Name Priya Srinivas
- Institute Rajiv Gandhi Centre for Biotechnology (RGCB)
Tool Details
- Tool name: MCF-7 sgRNA1 & 2 cell line
- Tool type: Cell Line
- Parental cell line: MCF7
- Organism: Human
- Tissue: Breast
- Gender: Female
- Cancer type: Breast Carcinoma
- Disease: Cancer
- Morphology: Epithelial slightly elongated morphology
- Growth properties: Adherent
- Model: Eipgenetic modification
- Model description: BRCA1 hypermethylated cells developed using CRISPR-Cas9 technology
- CRISPR: yes
- Conditional: yes
- Conditional description: conditional BRCA1 Hypermethylated cell line which gets reverted in the presence of tetracycline/doxycycline
- Receptors of note: Expression of BRCA1 only in the presence of doxycycline
- Application: For studying the BRCA1 non mutated and hypermethylated breast cancer tumorigenesis and drug development
- Description: The cells contain site-specific methylations in the BRCA1 promoter, which are analogous to the sites of methylation in breast cancer patients. Since, the direct effect of BRCA1 promoter hypermethylation in diagnosis and prognosis of breast and ovarian tumors has been largely obscure because of lack of proper in vitro and in vivo models these cell lines could be of potential research interest to others
- Research area: Cancer
- Production details: Cells with wildtype unmethylated BRCA1 background were infected with lentiviruses that encode for deadCas9- DNMT3A fusion protein and synthetic guide RNAs under the control of a tet-off promoter. After successful infection, the cells were then established following FACS sorting.
- Additional notes: 1?g/ml of doxycycline for 12 hours can stop the eGFP signal from pCL-CTIG and BRCA1 hypermethylation will be reverted.
- For Research Use Only
Target Details
Application Details
- Application: For studying the BRCA1 non mutated and hypermethylated breast cancer tumorigenesis and drug development
- Application notes: Pen-Strep, Invitrogen, Cat No. 15140125
Handling
- Format: Frozen
- Passage number: P3 (Post Generation)
- Growth medium: DMEM medium supplemented with 10% FBS and 1% Penicillin-Streptomycin
- Temperature: 37°C
- Atmosphere: 5% CO2
- Storage medium: 90% FBS + 1% Penicillin-Streptomycin + 10% sterile DMSO
- Storage conditions: Liquid Nitrogen
- Shipping conditions: Dry Ice
- Characterisation tests: RT-PCR, Western Blotting to confirm the expression of BRCA4
- Initial handling information: Store in Liquid Nitrogen. Thaw in 10% DMEM with 1% Penicillin-Streptomycin
- Subculture routine: Split the cells into 1:3 ratio when the cells reach 80% Confluency
- Cultured in antibiotics?: Yes
- Biosafety level: BSL-2
Documentation
- Available on request
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