The CRISPRaTest™ Functional dCas9-Activator Assay Kit (Cat.# CRATEST) measures functional dCas9 transactivator (e.g., dCas-VPH) in any mammalian cell system. The CRISPR gene activation system (CRISPRa), a modification of the original CRISPR knockout system, uses an sgRNA molecule to direct an inactive Cas9 (dCas9) fused with a gene activator, for example VP64 (i.e., dCas9-VP64), to a target location on genomic DNA near a gene promoter. The sgRNA-dCas9-Activator complex then increases expression levels of the gene regulated by the promoter.
The CRISPRaTest Kit provides reagents to measure the activator strength of any Streptococcus pyogenes dCas9 transactivator (dCas9-TA) hybrid protein in cells designed to be used for CRISPRa experiments using a FACS-based assay. The kit contains pseudoviral packaged lentiviral constructs with a green fluorescent protein (GFP) gene that increases in expression when expressed in cells with active dCas9-TA. The kit provides enough reagents to measure this activity in 5-15 cell lines.
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Please read the entire user manual before proceeding with your experiment. Also, please note that, when working with pseudoviral particles, you should follow the recommended guidelines for working with Biosafety Level 2 (BSL-2) materials.
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