• USA Home
  • CAS9D10AGFPP - CMV-CAS9D10A-2A-GFP Plasmid

EMAIL THIS PAGE TO A FRIEND
CAS9D10AGFPP Sigma-Aldrich

CMV-CAS9D10A-2A-GFP Plasmid

Synonym: CAS9D10A Plasmid

  •  NACRES NA.51

Purchase

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)
Promoter   Promoter name: CMV
reporter gene   GFP
selection   kanamycin
shipped in   dry ice
storage temp.   −20°C

Description

General description

The Cas9-D10A nickase plasmid co-expressing GFP uses the CMV promoter for strong transient expression of Cas9-D10A. Alternate promoters can be substituted by replacement of CMV using MluI and NheI. Also, the Cas9D10A expression plasmids can be linearized using XbaI for T7-based mRNA production.

Application

CMV-CAS9D10A-2A-GFP Plasmid has been used in CRISPR-Cas9 mutagenesis to truncate the forkhead box O3 (FOXO3) gene in the mammalian cell.

Functional Genomics
Use of Paired Cas9 Nickase + GFP for:

• Creation of gene knockouts in multiple cell lines
• Complete knockout of genes not amenable to RNAi
• Creation of knock-in cell lines with promoters, fusion tags or reporters integrated into endogenous genes

Components

1 vial containing 1ug of Cas9-D10A Nickase-GFP plasmid.

Please note, this product does not contain any guide RNA sequence. A pair of gRNA plasmids must be purchased separately through the Custom CRISPR paired nickase product tab.

Physical form

1 ug of Sigma Cas9-D10A nickase-GFP plasmid

Other Notes

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). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB. GFP is co-expressed from the same mRNA as the Cas9 nickase protein via a 2A peptide linkage, enabling tracking of transfection efficiency and enrichment of genome editing activity in cell populations via fluorescence activated cell sorting (FACS).

Legal Information

CRISPR Use License Agreement

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

CRISPR/Cas Nuclease RNA-guided Genome Editing

What is CRISPR/Cas9? How does CRISPR/Cas work? How does CRISPR Cas9 work? What is CRISPR/Cas9? Our role in developing CRISPR/Cas9
Keywords: Acetylations, Catalysis, Cell culture, Degradations, Gene expression, Genetic, PAGE, Polymorphisms, Transcription, Transduction, Transfection

Related Content

CRISPR

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...
Keywords: Genetic, Genomics, Transduction, Transfection

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

Peer-Reviewed Papers
15

References

Related Products

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?