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CRISPR21V

Sigma-Aldrich

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

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form

liquid

Quality Level

packaging

vial of 200 μL

concentration

(1x106 TU/ml (via p24 titering assay))

application(s)

CRISPR

shipped in

dry ice

storage temp.

−70°C

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CRISPR21SHC001HSHC203V
storage temp.

−70°C

storage temp.

−20°C

storage temp.

−70°C

storage temp.

−70°C

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

Quality Level

200

Quality Level

200

Quality Level

100, 200

Quality Level

100, 200

form

liquid

form

liquid

form

-

form

-

packaging

vial of 200 μL

packaging

vial of 50 μL

packaging

-

packaging

-

General description

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.

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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Articles

Genome-wide loss-of-function screening is a powerful approach to discover genes and pathways that underlie biological processes. Now complete knockout is achievable with two optimized gRNAs per gene. Minimized clone number ensures the most specific screening possible while controlling time and cost.

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