Components
Each kit consists of:
• one vial of Cas9 recombinant protein
• one vial containing 1 mL of 1× dilution buffer
• one vial containing 1 mL of nuclease-free water with glycerol
Packaging
pkg of 50 μg
pkg of 250 μg
Reconstitution
Lyophilized S. pyogenes Cas9 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
Recombinant Cas9 protein from Streptococcus pyogenes (~160 KD) is a ready-to-use reagent for genome engineering experiments. When combined with target-specific guide RNAs, wild type Streptococcus pyogenes Cas9 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.
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