CAS9GFPPRO

Cas9-GFP Protein

from Streptococcus pyogenes, fused with enhanced GFP, recombinant, expressed in E. coli, 3X NLS

• Synonym(s): Cas9-EGFP, SpCas9-EGFP, SpCas9-GFP
• UNSPSC Code: 12352202

Properties

• recombinant: expressed in E. coli
• Assay: ≥90% (SDS-PAGE)
• form: lyophilized powder
• packaging: pkg of 1 kit (3 components)
• reporter gene: GFP
• shipped in: wet ice
• storage temp.: −20°C

Description

General description

Recombinant Cas9-GFP protein from Streptococcus pyogenes (~194 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, wild type Streptococcus pyogenes Cas9-GFP 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.
An N-terminally fused enhanced green fluorescent protein with an excitation peak at 488 nm and emission peak at 509 nm allows for visualization of transfected RNP complex in addition to utility in flow cytometry applications.  The protein also contains three varied nuclear localization sequences postioned for optimal activity.

Application

• Functional Genomics
• Target Validation
• Genome Editing
• Fluorescence Microscopy
• Flow Cytometry

Features and Benefits

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

Packaging

• pkg of 50 μg( ≥260 pmol )
• pkg of 250 μg( ≥1300 pmol )

Components

Each kit consists of

• one vial of lyophilzed Cas9-GFP 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-GFP 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: 11 - Combustible Solids
• WGK: WGK 3