Sigma CRISPR dCAS9-p300V Activator Lenti Plasmid
SIGMA/DCAS9P300VP
Product Type: Chemical
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| application(s) | CRISPR |
| concentration | 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) |
| form | liquid |
| packaging | vial of 50 μL |
| Quality Level | 200  |
| recombinant | expressed in E. coli |
| shipped in | dry ice |
| storage temp. | −20°C |
| Application: | • Functional Genomics/Target Validation • Epigenetic Modification • Transcriptional Activation • Manufacture of dCas9-P300 expressing lentiviral particles |
| 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). Mutations to the catalytic domains, RuvC and HnH, render it inactive as a nuclease yet still allow for the protein to be programmed to target specific sequences of DNA with a single gRNA. A fusion of dCas9 to the catalytic histone acetyltransferase (HAT) core domain of the human E1A-associated protein p300 induces transcription by releasing DNA from its heterochromatin state allowing for continued and robust gene expression by endogenous cellular machinery. The dCas9-p300 CRISPR Gene Activator represents a distinct mechanism of action relative to dCas9-VP64 or other similar gene activation motifs |
| Features and Benefits: | The Sigma CRISPR dCas9p300V plasmid co-expresses p300-HAT and Blasticidin, allowing for blasticidin based selection of cells expressing dCas9p300. gRNAs can successfully direct nuclease-deficient Cas9 (dCas9) fused to p300 HAT catalytic domain to increase levels of histone acetylation and endogenous gene expression. The dCas9-p300 histone acetylation approach represents a distinct mechanism of action relative to dCas9-VP64 or other similar gene activation motifs. |
| General description: | This gene activation system is based on a fusion of inactive Cas9 (dCas9) to the catalytic histone acetyltransferase (HAT) core domain of the human E1A-associated protein p300. The dCas9-p300 activator lenti plasmids use the EF1 alpha promoter for strong expression of dCas9-P300 and blasticidin linked by a 2A peptide (EF1a-dCas9-P300-2A-Blast To order gRNA in any format click here |
| Legal Information: | CRISPR Use License Agreement Lentiviral, WPRE, and Evrogen Fluorophore License |
| 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 | 12352200 |