CRISPR/Cas9 Kinase Knockout Lentivirus Panel
Each CRISPR/Cas9 Kinase Knockout 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 (self-activation) 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.
The Custom Panel option allows you to pick between 2-50 kinases out of the full human kinase library. One vial of 200 µl of lentivirus is provided for each target. Download the table to view all available kinases.
Fill in the custom order with the list of kinases of interest (Gene names/symbols) and our customer service team will contact you shortly.
Example Flow Cytometry Data
Jurkat cells were transduced via spinoculation with ABL1 (panel A), ZAP70 (panel B) or ITK (panel C) CRISPR/Cas9 lentiviruses, at an MOI of approximately 1. Cells were selected using puromycin selection for 24-48 hours before antibody staining and flow cytometry analysis. Parental Jurkat cells are shown in green, and the transduced cells are shown in blue.
