All Photos(1)

CAS9RFPP

Sigma-Aldrich

CMV-CAS9-2A-RFP Plasmid

Synonym(s):
Cas9 plasmid
NACRES:
NA.51

Quality Level

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

RFP

shipped in

dry ice

storage temp.

−20°C

Looking for similar products? Visit Product Comparison Guide

General description

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. The addition of a fluorophore that is translationally co-expressed with the Cas9 nuclease allows for easy visualization of successful transfection.

Application

Functional Genomics/Target Validation
  • 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 1 μg of Cas9-2A-RFP plasmid.

Please note, product does not contain guideRNA sequence. This must be purchased separately through the Custom CRISPR product tab.

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). 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.

Physical form

Sigma Cas9-2A-RFP plasmid DNA is supplied at concentrations of 50ng/μl in 20μl.

Other Notes

Must be used in conjunction with a U6-gRNA plasmid in order to mediate a double strand break in the DNA.

Typical transfection concentrations used in literature are in the ranges of >= 1.0 μg/μL and <= 5 μL of Cas9 plasmid combined with >= 1.0 μg/μL and <= 5 μL of U6-gRNA plasmids. (All dosages above assume 0.5 to 1 million cells nucleofected).

Storage Class Code

12 - Non Combustible Liquids

WGK

nwg

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

Related Content

Gene Expression & Silencing

CRISPR, miRNA, siRNA, and shRNA technologies for gene expression and gene silencing applications.

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

Contact Technical Service