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To order a CRISPR/Cas9 lentivirus to knockout a kinase of interest, select the Gene Symbol of the kinase in the ordering window (for example, select AKT1 if ordering the lentivirus to knockout kinase AKT1). Download the table to view all available kinases.
The Kinase CRISPR/Cas9 lentivirus is designed to target a specific kinase of interest for knockout. The replication-incompetent, HIV-based, VSV-G pseudotyped lentiviral particles are ready to infect almost all types of mammalian cells, including primary and non-dividing cells. The SIN lentiviral backbone contains the Cas9 gene (Streptococcus pyogenes CRISPR associated protein 9) driven by an EF1A promoter, an sgRNA driven by a U6 promoter, and a puromycin selection marker. Each vial of lentivirus consists of a mixture of lentiviral particles targeting 5 different sgRNAs per gene.
The lentivirus integrates randomly into the cellular genome to express both Cas9 and the sgRNAs. Because it contains Cas9, the lentivirus can be used in any target cell regardless of whether the cells already express Cas9. Puromycin selection ensures high expression of both Cas9 and the sgRNAs. Knockout efficiencies will depend on the cell type and the gene of interest. Stable CRISPR-Cas9 knockout cell lines can also be generated following limiting dilution.
Figure 1: Schematic of the lentivector used to generate each kinase CRISPR/Cas9 Lentivirus.
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