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CRISPR16H

Sigma-Aldrich

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

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NACRES:
NA.51

form

liquid

Quality Level

packaging

vial of 8x25 μL (aliquots)

concentration

(5x108 TU/ml (via p24 titering assay))

application(s)

CRISPR

shipped in

dry ice

storage temp.

−70°C

Related Categories

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.

Application

Functional Genomics/Screening/Target Validation

Other Notes

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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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