CAS9RFPVP

Cas9-RFP Lenti Plasmid

• UNSPSC-Code: 41106609
• NACRES: NA.51

Eigenschaften

• Rekombinant: expressed in E. coli
• Form: liquid
• Verpackung: vial of 50 μL
• Konzentration: 20 ng/μL in TE buffer; DNA (1μg of Plasmid DNA)
• Anwendung(en): CRISPR
• Reportergen: RFP
• selection: ampicillin
• Versandbedingung: dry ice
• Lagertemp.: −20°C

Beschreibung

Allgemeine Beschreibung

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of Cas9 and RFP linked by a 2A peptide (EF1a-Cas9-2A-RFP) 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-RFP lenti plasmid is one part of a two part CRISPR system with individual Cas9 and gRNA expression vectors. For easy fluorescence microscopy a MOI greater than 200 should be employed when packaged into lentiviral particles.

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Anwendung

Functional Genomics/Target Validation

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

Leistungsmerkmale und Vorteile

• Highly specific
• Highly active
• Ready to use purified plasmid DNA

Prinzip

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.

Rechtliche Hinweise

CRISPR Use License Agreement

Lentiviral, WPRE and Evrogen Fluorophore Licenses

Sicherheitshinweise

• Lagerklassenschlüssel: 12 - Non Combustible Liquids
• WGK: WGK 2
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable