CRISPR dCas9-p300 RFP Activator Expression Plasmid

半岛bd体育手机客户端 说明

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 expression plasmids use the CMV promoter for strong transient expression of dCas9-P300 and RFP linked by a 2A peptide (CMV-dCas9-p300-2A-RFP) allowing for easy visualization of successful transfection. The dCas9-P300-RFP expression plasmid is one part of a two part CRISPR system with individual dCas9-P300 and gRNA expression vectors.

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应用

Functional Genomics/Target Validation

• Epigenetic Modification
• Transcriptional Activation
• Manufacture of dCas9-P300 expressing lentiviral particles

特点和优势

The Sigma CRISPR dCas9p300 plasmid co-expresses p300-HAT and RFP, to easily monitor delivery and expression in your target cell type. 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.

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 bacteriumStreptococcus pyogeneshas 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 catalystic domains, RuvC and HnH, render it inactive as a nuclease yet still allow for the protein to be programmed with a single gRNA, directing p300 epigenetic modification cargo to any desired genomic location.

法律信息

CRISPR Use License Agreement

Evrogen Fluorophore Licenses

半岛bd体育手机客户端 性质

质量水平	200
重组	expressed inE. coli
包装	vial of 50 μL
浓度	20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
application(s)	CRISPR
运输	dry ice
储存温度	?20℃

安全信息

储存分类代码	12 - Non Combustible Liquids
WGK	WGK 2
闪点(F)	Not applicable
闪点(C)	Not applicable