Cas9 Nuclease is the purified recombinant Streptococcus pyogenes Cas9enzyme containing a nuclear localization signal (NLS) at the C-terminalfor targeting to the nucleus. This enzyme is designed to performCRISPR/Cas9-mediated genome editing. The physical purity of thisenzyme is ≥98% as assessed by SDS-PAGE with Coomassie blue staining.

CRISPR (Clustered regularly interspaced short palindromic repeats)/CRISPR-associated (Cas) systems provide bacteria and archaeawith adaptive immunity against viruses and plasmids by using CRISPR RNAs(crRNAs) to guide the silencing of invading nucleic acids. TheCRISPR system consists of a short non-coding guide RNA (sgRNA) made upof a target complementary CRISPR RNA (crRNA) and an auxiliarytransactivating crRNA (tracrRNA). The sgRNA guides the Cas9 endonucleaseto a specific genomic locus via base pairing between the crRNA sequenceand the target sequence, and cleaves the DNA to create a double-strandbreak. The location of the break is within the target sequence 3 basesfrom the NGG PAM (Protospacer Adjacent Motif). The PAM sequence,NGG, must follow the targeted region on the opposite strand of the DNAwith respect to the region complementary sgRNA sequence.

Cas9 nuclease is free from detectable RNase, Endonuclease (nicking) and non-specific DNase activities.

Product Includes

• Cas9 Nuclease (160 ng/µl (1µM))
• 10x Cas9 Nuclease Reaction Buffer

Source: E. coli BL21 (DE3) strain expressing a Cas9 gene from Streptococcuspyogenes with an N-terminal 6xHis tag and C-terminal SV40 nuclearlocalization signal (NLS)

Storage/Handling: Store at -20°C. Product may be shipped on blue ice.