MDA MB-231 sgRNA1 & 2 cell line

Tool Details

*FOR RESEARCH USE ONLY (for other uses, please contact the licensing team)

• Name: MDA MB-231 sgRNA1 & 2 cell line
• Cancer: Breast cancer
• Cancers detailed: Breast carcinoma
• Research fields: Cancer
• Parental cell: MDA MB 234
• Organism: Human
• Gender: Female
• Tissue: Breast
• Donor: Established from the pleural effusion from a 69 year female caucasian suffering from a breast adenocarcinoma. Cells may carry B or C type retrovirus and are considered to represent a category 2 pathogen (P2 containment)
• 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
• 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
• Application: For studying the BRCA1 non mutated and hypermethylated breast cancer tumorigenesis and drug development
• 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.
• Biosafety level: 2
• Additional notes: 1?g/ml of doxycycline for 12 hours can stop the eGFP signal from pCL-CTIG and BRCA1 hypermethylation will be reverted.
• Recommended controls: MDA-MB-231 sgRNA Control cell line, MCF-7 sgRNA1 controls Control cell line , MCF-7 sgRNA2 Control cell line

Applications

• Application: For studying the BRCA1 non mutated and hypermethylated breast cancer tumorigenesis and drug development
• Application notes: Pen-Strep, Invitrogen, Cat No. 15140129

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
• Unit size: 1x10^6 cells / vial
• Shipping conditions: Dry Ice
• Storage medium: 90% FBS + 1% Penicillin-Streptomycin + 10% sterile DMSO
• Storage conditions: Liquid Nitrogen
• 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
• Characterisation tests: RT-PCR, Western Blotting to confirm the expression of BRCA8
• Str profiling: STR Profile confirmed on 13.01.2028

References

• https://doi.org/10.1101/2022.04.30.490089