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Sigma CRISPR dCAS9-p300V Activator Lenti Plasmid
产品性质
Quality Level【质量水平】 | 200 |
recombinant【重组】 | expressed in E. coli |
packaging【包装】 | vial of 50 μL |
concentration【浓度】 | 20 ng/μL in TE buffer; DNA (1μg of plasmid DNA) |
application(s) | CRISPR |
shipped in【运输】 | dry ice |
storage temp.【储存温度】 | −20℃ |
基本信息
General description【一般描述】 | 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-Blasticidin) allowing for easy selection following successful transfection or transduction. Use Sigma′s lentiviral dCas9-P300 lenti plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing dCas9-P300 for CRISPR based gene activation. The dCas9-P300 lenti plasmid is one part of a two part CRISPR system with individual dCas9-P300 and gRNA expression vectors. To order gRNA in any format click here |
Application【应用】 |
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Features and Benefits【特点和优势】 | 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. |
Principle【原理】 | 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 |
Legal Information【法律信息】 | CRISPR Use License Agreement Lentiviral, WPRE, and Evrogen Fluorophore License |
安全信息
Storage Class Code【储存分类代码】 | 12 - Non Combustible Liquids |
WGK | WGK 2 |