CAS9HYGROVP

Cas9 Hygromycin Lenti Plasmid

Properties

• recombinant: expressed in E. coli
• form: liquid
• packaging: vial of 50 μL
• concentration: 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
• application(s): CRISPR
• selection: blasticidin
• shipped in: dry ice
• storage temp.: −20°C

Description

General description

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of Cas9 and a hygromycin resistance cassette linked by a 2A peptide (EF1a-Cas9-2A-Hygro) allowing for easy visualization of successful transfection or transduction. Use Sigma′s lentiviral Cas9 plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing Cas9 for CRISPR based genome editing. Sigma′s Cas9-Hygro lenti plasmid is one part of a two part CRISPR system with individual Cas9 and gRNA expression vectors.

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Application

Functional Genomics/Target Validation

• Creation of gene knockouts in multiple cell lines
• Complete knockout of genes not amenable to RNAi
• Manufacture of Cas9 expressing Lentiviral Particles

Features and Benefits

• Highly specific and highly active
• Sequence verified
• Ready to use purified plasmid DNA

Principle

CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.

Legal Information

CRISPR Label License

Lentiviral, WPRE, and Evrogen Licenses

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable