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ESPCAS9GFP

Sigma-Aldrich

CMV-ESPCAS9-2A-GFP Plasmid

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recombinant

expressed in E. coli

Quality Level

packaging

vial of 50 μL

concentration

20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)

shipped in

dry ice

storage temp.

−20°C

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General description

This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of eSpCas9 and GFP linked by a 2A peptide (CMV-eSpCas9-2A-GFP) 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.

To order gRNA in any format click here

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.

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)

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