CMV-ESPCAS9-2A-RFP Plasmid
SIGMA/ESPCAS9RFP
Product Type: Chemical
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| concentration | 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) |
| packaging | vial of 50 μL |
| promoter | Promoter name: CMV |
| Quality Level | 200  |
| recombinant | expressed in E. coli |
| reporter gene | RFP |
| selection | kanamycin |
| shipped in | dry ice |
| storage temp. | −20°C |
| Application: | 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 |
| Biochem/physiol Actions: | 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. |
| Features and Benefits: | • Enhanced specificity compared to wild type Cas9 • Highly active • Ready to use purified plasmid DNA |
| General description: | 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 |
| Legal Information: | CRISPR Use License Agreement Evrogen Fluorophore Licenses |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 41106609 |