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ESPCAS9RFP

Sigma-Aldrich

CMV-ESPCAS9-2A-RFP Plasmid

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About This Item

Código UNSPSC:
41106609
NACRES:
NA.51

recombinante

expressed in E. coli

Nível de qualidade

embalagem

vial of 50 μL

concentração

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

Promotor

Promoter name: CMV

gene repórter

RFP

Seleção

kanamycin

Condições de expedição

dry ice

temperatura de armazenamento

−20°C

Descrição geral

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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Aplicação

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

Características e benefícios

  • Enhanced specificity compared to wild type Cas9
  • Highly active
  • Ready to use purified plasmid DNA

Princípio

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.

Código de classe de armazenamento

10 - Combustible liquids

Classe de risco de água (WGK)

WGK 2

Ponto de fulgor (°F)

Not applicable

Ponto de fulgor (°C)

Not applicable


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Certificados de análise (COA)

Lot/Batch Number

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

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