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DCAS9P300 Sigma-Aldrich

Sigma CRISPR dCas9-p300 Activator Expression Plasmid

  •  NACRES NA.51

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Properties

Related Categories CRISPR-Cas9, Cas9-only (DNA, mRNA, and virus), Functional Genomics and RNAi, Molecular Biology
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

A gene activation system based on a fusion of inactive Cas9 or dCas9 to the catalytic histone acetyltransferase (HAT) core domain of the human E1A-associated protein p300. The dCas9-p300 CRISPR Gene Activator with the p300 domain represents a distinct mechanism of action relative to dCas9-VP64 or other similar gene activation motifs. The p300 histone acetyltransferase protein opens a transcriptional highway by releasing the DNA from its heterochromatin state and allowing for continued and robust gene expression by the endogenous cellular machinery. The Cas9 expression plasmids use the CMV promoter for strong transient expression of Cas9. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9 expression plasmids can be linearized using XbaI for T7-based mRNA production.

Application

• Functional Genomics/Target ValidationEpigenetic Modification
• Transcriptional Activation

Features and Benefits

• The Sigma CRISPR dCas9p300 plasmid co-expresses p300-HAT and GFP, to easily monitor delivery and expression in your target cell type (figure 1)gRNAs can successfully direct nuclease-deficient Cas9 (dCas9) fused to p300 HAT catalytic domain to increase levels of histone acetylation and endogenous gene expressionThe dCas9-p300 histone acetylation approach represents a distinct mechanism of action relative to dCas9-VP64 or other similar gene activation motifs

Components

1 vial containing 1ug of dCas9-p300 plasmid.

Linkage

To order U6-gRNA plasmids, please visit sigma.com/crisprs


Click here for more information on dcas9-p300 Gene Activator Expression Plasmid.

Other Notes

Must be used with U6-gRNA only expressing plasmid.

Safety & Documentation

Safety Information

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

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

CRISPR-based Gene Activation

Epigenome editing is a tool in which the DNA or histone is modified at specific sites in the genome using engineered molecules. This strategy requires precise targeting which is accomplished through ...
Keywords: Acetylations, Cancer, Catalysis, Gene expression, Transcription

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In recent years CRISPR has revolutionized gene editing capabilities, leading to sophisticated ways to create success with any experiment. As the first company to offer custom biomolecules globally fo...
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Predictive Models for Neuroscience using CRISPR [VIDEO]

Caroline Beckett, the global CRISPR product manager, discusses reagent solutions for creating predictive models for neuroscience research. She notes that many neuroscientists want to be able to creat...
Keywords: Cell culture, Diseases, Neurodegenerative Diseases, Neuroscience

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