Lenti CRISPR Human HPRT1 Positive Control Transduction Particles (LV04 vector)

The Lenti CRISPR Human HPRT1 Positive Control Transduction Particles (LV04 vector) product is a critical positive control to monitor transduction efficiency for lentiCRISPRs. This control is produced from the sequence-verified CRISPR lentiviral plasmid targeting human HPRT1. 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 HPRT1 positive control uses 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.

Lentiviral-based particles permit efficient infection and integration of the construct into differentiated and non-dividing cells, such as neurons and dendritic cells, overcoming low transfection and integration difficulties when using these cell lines. Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging and envelope plasmids.

Functional Genomics/Screening/Target Validation

Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells, respectively.