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ESPCAS9BST

Sigma-Aldrich

eSpCas9-Blasticidin Lenti 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)

selection

ampicillin

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of eSpCas9 and a blasticidin resistance cassette linked by a 2A peptide (EF1a-eSpCas9-2A-Blasticidin) allowing for easy selection following successful transfection or transduction. Use Sigma′s lentiviral eSpCas9 plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing eSpCas9 for CRISPR based genome editing. Sigma′s lenti-eSpCas9 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
  • Manufacture of eSpCas9 expressing Lentiviral Particles

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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Customers Also Viewed

Rationally engineered Cas9 nucleases with improved specificity.
Slaymaker, I.M., et al.
Science, 351, 84-88 (2015)

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