CRISPR13V
CRISPR-Lenti Human HPRT1 Positive Control Transduction Particles
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About This Item
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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.
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
To see more application data, protocols, vector maps visit sigma.com/crispr.
- 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
Storage Class
12 - Non Combustible Liquids
wgk_germany
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
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