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CRISPR16H Sigma-Aldrich

Lenti CRISPR HPRT1 gRNA Control Transduction Particles, High Titer (LV02 vector)

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Properties

Related Categories CRISPR Controls (DNA and Virus), CRISPR-Cas9, Functional Genomics and RNAi, Molecular Biology
packaging   vial of 8x25 μL (aliquots)
concentration   (5x108 TU/ml (via p24 titering assay))
application(s)   CRISPR: suitable
shipped in   dry ice
storage temp.   −70°C

Description

Application

Functional Genomics/Screening/Target Validation

General description

The Lenti CRISPR HPRT1 gRNA Control Transduction Particles (LV02 vector) product is a critical positive control used to monitor transduction 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 uses a dual-component system consisting of a U6-driven guide RNA plasmid targeting the human HPRT1 gene and Ef1a-driven Puromycin resistance casette. 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.

Legal Information

CRISPR Use License Agreement

Lentiviral, WPRE, and Evrogen Licenses

Other Notes

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

Safety & Documentation

Safety Information

RIDADR 
UN 3245 9

Documents

Certificate of Analysis

Protocols & Articles

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