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recombinant
expressed in E. coli
form
liquid
packaging
vial of 50 μL
concentration
20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
application(s)
CRISPR
promoter
Promoter name: CMV
selection
kanamycin
shipped in
dry ice
storage temp.
−20°C
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.
To order gRNA in any format click here
To order gRNA in any format click here
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
Storage Class
12 - Non Combustible Liquids
wgk_germany
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
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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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