Cas9 Nuclease, S. pyogenes, is an RNA-guided endonuclease that catalyzes site-specific cleavage of double stranded DNA. The location of the break is within the target sequence 3 bases from the NGG PAM (Protospacer Adjacent Motif) (1). The PAM sequence, NGG, must follow the targeted region on the opposite strand of the DNA with respect to the region complementary sgRNA sequence.Cas9 in vivo: Bacterial Adaptive ImmunityCRISPR loci in the bacterial genome are transcribed and processed into crRNA. Cas9 endonuclease complexed with a crRNA and separate tracrRNA cleaves foreign DNA containing a 20-nucleotide crRNA complementary sequence adjacent to the PAM sequence. (Figure not drawn to scale.)Cas9 nuclease, S. pyogenes, complexed with an sgRNACleavage occurs three nucleotides upstream of the PAM sequence (shown in red). sgRNAs are complimentary to the strand of DNA opposite of the PAM.In vitro DNA cleavage using Cas9 Nuclease, S. pyogenesReactions were set up according to recommended conditions, and cleavage products were resolved on a 1% agarose TBE gel. Input DNA is PvuII-linearized pBR322 plasmid DNA.
Product Source
A His-tagged engineered variant of E. coli that carries the cloned Cas9 gene from Streptococcus pyogenes.
This product is related to the following categories:
Genome Editing Products,
Programmable Nucleases
This product can be used in the following applications: