CRISPR-associated endonuclease Cas9 (UniProt: J7RUA5; also known as SaCas9) is encoded by the Cas9 gene (Gene ID: in Staphylococcus aureus. CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer.Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes the protospacer adjacent motif (PAM) in the CRISPR repeat sequences to help distinguish self-versus non-self, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity. Cas9 endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, and then trimmed 3'-5' exonucleolytically.
Synonyms: SaCas9, CRISPR-associated endonuclease Cas-9
Application: Western Blotting Analysis: 0.57 µg/mL from a representative lot detected CRISPR-associated endonuclease Cas9/Csn1 in whole cell lysates of HEK293 (WT), HEK293/Cas9 from S. aureus, or HeLa/Cas9 from S. pyogenes. (Courtesy of Stefan Schuchner, Ph.D. and Egon Ogris, M.D., Medical University of Vienna, Austria).Immunofluorescence Analysis: A 1:1,000 dilution from a representative lot detected CRISPR-associated endonuclease Cas9/Csn1 in transiently transfected U20S cells expressing SaCas9-T2A-GFP. (Courtesy of Stefan Schuchner, Ph.D. and Egon Ogris, M.D., Medical University of Vienna, Austria).Immunoprecipitation Analysis: 1-2 µg from a representative lot detected CRISPR-associated endonuclease Cas9/Csn1 in SaCas9. (Courtesy of Stefan Schuchner, Ph.D. and Egon Ogris, M.D., Medical University of Vienna, Austria).
Other Notes: Concentration: Please refer to lot specific datasheet.