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CRISPR13V

Sigma-Aldrich

CRISPR-Lenti Human HPRT1 Positive Control Transduction Particles

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About This Item

UNSPSC Code:
41106609
NACRES:
NA.51

form

liquid

Quality Level

packaging

vial of 200 μL

concentration

1x106  VP/ml (via p24 assay)

application(s)

CRISPR

shipped in

dry ice

storage temp.

−70°C

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.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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Protocols

Learn about CRISPR Cas9, what it is and how it works. CRISPR is a new, affordable genome editing tool enabling access to genome editing for all.

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