Cas9-GFP Protein

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 GenomicsTarget ValidationGenome EditingFluorescence MicroscopyFlow Cytometry
• Features and Benefits: Highly specificHighly activeReady-to-inject/transfect
• Packaging: pkg of 50 μg( ≥260 pmol )pkg of 250 μg( ≥1300 pmol )
• Components: Each kit consists ofone vial of lyophilzed Cas9-GFP recombinant proteinone vial containing 1 mL of 1× dilution bufferone 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 gRNACheck out our other MISSION® Cas9 Proteins at SigmaAldrich.com/CRISPRproteins
• Legal Information: CRISPR Use License Agreement