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DCAS9P300RFP Sigma-Aldrich

CRISPR dCas9-p300 RFP Activator Expression Plasmid

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Properties

Quality Level   200
recombinant   expressed in E. coli
packaging   vial of 50 μL
concentration   20 ng/μL in TE buffer; DNA (1μg of plasmid DNA)
application(s)   CRISPR: suitable
shipped in   dry ice
storage temp.   −20°C

Description

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 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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Application

Functional Genomics/Target Validation
• Epigenetic Modification
• Transcriptional Activation
• Manufacture of dCas9-P300 expressing lentiviral particles

Features and Benefits

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.

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). 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.

Legal Information

CRISPR Use License Agreement

Evrogen Fluorophore Licenses

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 2
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

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Protocols & Articles

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