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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 transcriptional activator VP64. Stable dCas9-VP64 expression is maintained with Blasticidin resistance. Cells also stably express P65 (Transcription Factor p65, or Nuclear Factor NF-κB p65) and HSF1 (Heat Shock Factor 1) fused with an MS2 tag, which is maintained with Hygromycin resistance. When these cells are transfected with an MS2- tagged sgRNA targeting the promoter region of the gene of interest, dCas9-VP64 and MS2-P65-HSF1 are recruited to the genomic DNA and begin transcription, inducing expression of the desired gene.
Host Cell Line: HeLa
Mycoplasma Testing: This cell line has been screened using the MycoAlert™ Mycoplasma Detection Kit (Lonza, #LT07-118) to confirm the absence of Mycoplasma contamination. MycoAlert Assay Control Set (Lonza, #LT07-518) was used as a positive control.
Storage Stability: Cells will arrive upon dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt.
Supplied As: Each vial contains 2 x 106 cells in 1 ml of 10% DMSO
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: Konermann, S., et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 2015. 517(7536): 583-588.
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