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ESPCAS9RFP

Sigma-Aldrich

CMV-ESPCAS9-2A-RFP Plasmid

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About This Item

UNSPSC-Code:
41106609
NACRES:
NA.51

Rekombinant

expressed in E. coli

Qualitätsniveau

Verpackung

vial of 50 μL

Konzentration

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

Promoter

Promoter name: CMV

Reportergen

RFP

selection

kanamycin

Versandbedingung

dry ice

Lagertemp.

−20°C

Allgemeine Beschreibung

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.

To order gRNA in any format click here

Anwendung

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

Leistungsmerkmale und Vorteile

  • Enhanced specificity compared to wild type Cas9
  • 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. 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.

Lagerklassenschlüssel

10 - Combustible liquids

WGK

WGK 2

Flammpunkt (°F)

Not applicable

Flammpunkt (°C)

Not applicable


Analysenzertifikate (COA)

Suchen Sie nach Analysenzertifikate (COA), indem Sie die Lot-/Chargennummer des Produkts eingeben. Lot- und Chargennummern sind auf dem Produktetikett hinter den Wörtern ‘Lot’ oder ‘Batch’ (Lot oder Charge) zu finden.

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In der Dokumentenbibliothek finden Sie die Dokumentation zu den Produkten, die Sie kürzlich erworben haben.

Die Dokumentenbibliothek aufrufen

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

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