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Applications: 1. Quickly generating CRISPR-activated cell pools or cell lines in Jurkat cells.
2. Implementing sgRNA CRISPRa screens in Jurkat 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 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.
Host Cell Line: Jurkat
Storage Stability: Store in liquid nitrogen immediately upon receipt.
Supplied As: Each vial contains ~ 2 x 10^6 cells in 1 ml of 10% DMSO in FBS.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-1
References: Konermann, S., et al. (2015). Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 517(7536): 583-588.
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