All Photos(1)

CAS9D10APR

Sigma-Aldrich

Cas9-D10A Nickase Protein

from Streptococcus pyogenes with D10A mutation, recombinant, expressed in E. coli, 1X NLS

Synonym(s):
Cas9 Nickase, Nickase Cas9, Cas9D10A
NACRES:
NA.51

Quality Level

assay

≥95% (SDS-PAGE)

form

lyophilized powder

packaging

pkg of 1 kit (3 components)

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

Recombinant Cas9-D10A Nickase protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific paired guide RNAs, S. pyogenes Cas9-D10A nickase will act as a targeted nuclease suitable for transfection of cell cultures and for the accelerated development of genetically-modified animals via one-cell embryo injection.
Cas9 (CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein-9 nuclease) nickase creates double-stranded breaks through two nuclease domains, named RuvC and HNH.

Application

Functional Genomics/Target Validation/Genome Editing

Features and Benefits

  • Highly specific
  • Highly active
  • Ready-to-inject/transfect

Packaging

pkg of 50 μg (≥ 300 pmol)
pkg of 250 μg (≥ 1500 pmol)

Components

Each kit consists of:
  • one vial of Cas9-D10A Nickase recombinant protein
  • one vial containing 1 mL of 1× dilution buffer
  • one vial containing 1 mL of nuclease-free water with glycerol

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 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.

Reconstitution

Lyophilized S. pyogenes Cas9-D10A Nickase protein should be resuspended in the Reconstitution solution provided to desired concentration. Gently tap tube to completely dissolve lyophilized powder, incubate for 10 minutes on ice, and spin tube to bring material to bottom of tube.

Other Notes

Use our CRISPR Selection Tool to order gRNA

Check out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins

See the product in our recent article on bioRxiv Highly efficient and specific genome editing in human cells with paired CRISPR-Cas9 nickase ribonucleoproteins

Legal Information

MISSION is a registered trademark of Sigma-Aldrich Co. LLC

Storage Class Code

11 - Combustible Solids

Certificate of Analysis

Certificate of Origin

Stem Cell Genetics for Biomedical Research: Past, Present, and Future, 376-376 (2018)

Protocols

Ribonucleoprotein (RNP) Protocols using SygRNA ® gRNAs and Cas9 proteins

Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.

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