The Lenti CRISPR Human HPRT1 Positive Control Plasmid DNA (LV04 vector) is a critical positive control to monitor transfection efficiency for lentiCRISPRs. The protein encoded by this gene is a transferase which plays a central role in the generation of purine nucleotides. Cells with intact HPRT1 will die upon exposure to 6-TG (Product No. A4882), an antimetabolite used in the treatment of leukemias. Conversely, cells with HPRT1 knocked out will increase over time with 6-TG selection. This targets a validated HPRT1 CRISPR site, which serves as an experimental control for lenti CRISPRs. The lenti CRISPR positive control is a dual-component system consisting of a U6-driven guide RNA plasmid targeting the human HPRT1 gene and hPGK-driven Puromycin and BFP expression. This control requires Cas9 be delivered by a separate vector.
Application
Functional Genomics/Screening/Target Validation
Other Notes
Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells, respectively. Self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids (Product No. SHP002).
Genome-wide screening with optimized gRNAs per gene ensures specific and efficient knockout, controlling time and cost.
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