ESPCAS9RFP

CMV-ESPCAS9-2A-RFP Plasmid

• UNSPSC Code: 12352200

Properties

• recombinant: expressed in E. coli
• Quality Level: 200
• packaging: vial of 50 μL
• concentration: 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
• Promoter: Promoter name: CMV
• reporter gene: RFP
• selection: kanamycin
• shipped in: dry ice
• storage temp.: −20°C

Description

General description

This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of eSpCas9 and RFP linked by a 2A peptide (CMV-eSpCas9-2A-RFP) allowing for easy visualization of successful transfection. The eSpCas9 expression plasmid is one part of a two part CRISPR system with individual eSpCas9 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
• Creation of knock-in cell lines with promoters, fusion tags, or reporters integrated into endogenous genes

Features and Benefits

• Enhanced specificity compared to wild type Cas9
• Highly active
• 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. Newly engineered eSpCas9 enables the efficient targeted gene editing of established CRISPR systems with the benefit of reduced off-target effects. Point mutations in the chromosome-binding motif of SpCas9, as described by Slaymaker, et al., provide higher on-target fidelity without loss of cleavage efficiency.

Legal Information

CRISPR Use License Agreement

Evrogen Fluorophore Licenses

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable