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CAS9D10APR Sigma-Aldrich

Cas9-D10A Nickase Protein

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

Synonym: Cas9 Nickase, Cas9D10A, Nickase Cas9

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Properties

Related Categories CRISPR-Cas9, Functional Genomics and RNAi, MISSION™ Cas9 Proteins, Molecular Biology
Quality Level   PREMIUM
assay   ≥95% (SDS-PAGE)
form   lyophilized powder
storage temp.   −20°C

Description

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

Other Notes

Use our CRISPR Selection Tool to order gRNA

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

Packaging

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

Legal Information

CRISPR Use License Agreement

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

Reconstitution

Lyophilized S. pyogenes Cas9-D10A Nickase protein should be resuspended in the Reconstitution solution provided to desired concentration. The solution should be allowed to incubate at room temperature (20-23 °C) for 10 minutes. Centrifuge immediately before use.

Application

Functional Genomics/Target Validation/Genome Editing

Features and Benefits

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

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.

General description

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.

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.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Spring into Genome Editing
Protocols & Articles

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Peer-Reviewed Papers
15

References

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