Application

Functional Genomics/Screening/Target Validation Positive control, HPRT1, CRISPR lentivirus format, for creating gene knockouts/genetic modifications in cell lines.To see more application data, protocols, vector maps visit sigma.com/crispr.

Features and Benefits

Serves as a positive experimental control for the CRISPR editing workflow using lentiCRISPRs (virus format). Allows for validation of your system with the lentiCRISPR/Cas9 system. A positive result in a mis-match detection assay will indicate validation of your system.

General description

The Sigma CRISPR-LENTI HUMAN HPRT1 POSITIVE CONTROL TRANSDUCTION PARTICLES are 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 (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 Sigma lenti CRISPRs. The Sigma lenti CRISPR positive control is a one-component system consisting of an EF1-alpha- driven Cas9, a U6-driven guide RNA plasmid targeting the human HPRT1 gene, with both Puromycin and GFP co-expressed with Cas9.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 plasmids.

Legal Information

CRISPR Use License AgreementLentiviral and WPRE License Agreements

Physical form

200 µl of 10⁶ TU/ml (via p24 titering assay) lentiviral particles are provided as frozen stock.