CRISPR13V

CRISPR-Lenti Human HPRT1 Positive Control Transduction Particles

• UNSPSC Code: 12352200
• NACRES: NA.51

Properties

• form: liquid
• Quality Level: 200
• packaging: vial of 200 μL
• concentration: 1x10⁶ VP/ml (via p24 assay)
• application(s): CRISPR
• shipped in: dry ice
• storage temp.: −70°C

Description

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.

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.

Physical form

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

Legal Information

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 3
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable