recombinant

expressed in E. coli

packaging

vial of 50 μL

concentration

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

application(s)

CRISPR: suitable

Promoter

Promoter name: CMV

selection

kanamycin

shipped in

dry ice

storage temp.

−20°C

Verwandte Kategorien

General description

This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of Dead Cas9 (CMV-dCas9). The dCas9 expression plasmid is one part of a two part CRISPR system with individual dCas9 and gRNA expression vectors.

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Application

Functional Genomics/Target Validation
  • Proxy CRISPR
  • Suitable for genomic DNA detection

Features and Benefits

  • Highly specific
  • Sequence verified
  • 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 rendered inactive (dCas9) with mutations to the two protein domains, RuvC and HnH (D10A and H840A respectively), which are responsible for nuclease activity. The nuclease deficient protein can then be programmed with a gRNA and directed to bind at a desired sequence of DNA. Variants of programmable endonucleases are often unable to cleave a large number of the targets that are efficiently cleaved by canonical SpCas9 in human cells. Cotargeting dead Cas9 may alter local chromatin structures and render otherwise inaccessible target sites now accessible and cleavable by alternative nucleases such as type II-B FnCas9, type II-C CjCas9 and NcCas9 and type V FnCpf1.

Legal Information

RIDADR

NONH for all modes of transport

WGK Germany

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Targeted activation of diverse CRISPR-Cas systems for mammalian genome editing via proximal CRISPR targeting.
Chen, F. et al.
Nature Communications, 8 (2017)

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