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Applications: Quickly generate CRISPR-activated cell pools or cell lines in MDA-MB-231 cells.Implement sgRNA CRISPRa screens in MDA-MB-231 cells.
Description: This cell line has been engineered for use with the CRISPR Synergistic Activation Mediator (SAM) system to induce transcriptional activation and expression of any gene of interest. Cells stably express a mutated dCas9 (Streptococcus pyogenes CRISPR-associated protein 9), lacking any endonuclease activity, fused to VP64, a transcriptional activator. Stable dCas9-VP64 expression is maintained with Blasticidin resistance. Cells also stably express a construct consisting of P65 (Transcription Factor p65, or Nuclear Factor NF-κB p65) and HSF1 (Heat Shock Factor 1) fused with an MS2 tag (from the MS2 coat protein), which is maintained with Hygromycin resistance. When these cells are transfected with an MS2 aptamer fused to a single guide RNA (sgRNA)targeting the promoter region of the gene of interest, dCas9-VP64 and MS2-P65-HSF1 are recruited to the promoter in the genomic DNA by the MS2 aptamer and begin transcription, inducing high level expression of the desired protein.Cells were functionally validated by transfecting the cells with an sgRNA-MS2 targeting PD-1, resulting in high expression of PD-1.
Host Cell Line: MDA-MB-231 cells, human breast mammary gland cells, epithelial, adherent
Mycoplasma Testing: The cell line has been screened to confirm the absence of Mycoplasma species.
Storage Stability: Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.
Supplied As: Each vial contains 2 x 106 cells in 1 ml of cell freezing medium (BPS Bioscience #79796)
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-1
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