CRISPRa (SAM) HepG2 Cell Line

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 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 site in the genomic DNA and begin transcription, inducing expression of the desired gene.

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