CAS9PL

Cas9 Plus Protein

from Streptococcus pyogenes, recombinant, expressed in E. coli, 3X NLS

• Synonym(s): Cas9, SpCas9, SpyCas9

Properties

• recombinant: expressed in E. coli
• Quality Level: 200
• Assay: ≥95% (SDS-PAGE)
• form: lyophilized powder
• packaging: pkg of 1 kit (4 components)
• application(s): CRISPR
• shipped in: wet ice
• storage temp.: −20°C

Description

General description

Recombinant Cas9 Plus protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, Streptococcus pyogenes Cas9 Plus protein 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.

Application

Functional Genomics; Target Validation; Genome Editing

Features and Benefits

• Enhanced specificity compared to wild type Cas9
• Increased activity compared to enhanced specificity Cas9
• 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 Plus Recombinant Protein
• one vial of Plus Transfection Buffer
• one vial of Reconstitution Buffer
• one vial of Dilution Buffer

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 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

Legal Information

CRISPR Use License Agreement

MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 3