Description
CRISPRtest™ Essential-Gene Cas9 Activity Kit is an easy-to-use, highly sensitive kit for measuring functional Cas9 activity in human or mouse cell lines. Note, because of the way the Essential Gene Cas9 Assay works, there are different kits for Human and Mouse cells. The mouse cell kits will not work with human cells, nor the human kits work with mouse cells. Be sure to pick the correct kit for your cell type with the fluorescent marker combination that will work in your cells. The Essential Gene Cas9 Assays contain two pseudoviral-packaged, lentiviral constructs--one (1) that expresses an sgRNA that targets an essential gene and one fluorescent protein (e.g., GFP, BFP) as well as a second construct with a non-target sgRNA and a second fluorescent marker (usually RFP or BFP). After transduction, knockout of the essential target gene kills off the cells with the one fluorescent marker. Measurement of the depletion of the one fluorescence marker due to the death of cells with the lethal sgRNA as compared to cells harboring the fluorescent marker with the non-targeting sgRNA provides a quantitative estimate of Cas9 activity. This Essential Gene Cas9 Assays work differently than Cellecta's Cas9 Fluorescent Reporter Assay (i.e., CRISPRuTest, CRISPRiTest, and CRISPRaTest kits). While readout for both types of Cas9 assays relies on the the disappearance of a fluorescent marker to assess Cas9 activity, the Essential-Gene Cas9 relies on cell death induced by disruption of a gene required for viability, whereas, with the CRISPR-test Fluorescent Reporter Assays, the sgRNA directly interferes with expression of one of the reporter genes. Each kit provides enough reagents to measure this activity in 5-10 cell lines