CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) belongs to the type II CRISPR/CAS9 system. It is part of an adaptive immune system of the Streptococcus pyogenes SF370, protecting from pathogens targets genes by cleaving the foreign DNA in a sequence-dependent manner.2 The type II CRISPR/Cas system which has been adapted to expression in eukaryotic cells, consists of four genes including the Cas9 (CRISPR-associated proteins) nuclease, two noncoding CRISPR RNAs (crRNAs, or gRNA), trans-activating crRNA (tracrRNA) and a precursor crRNA (pre-crRNA) array. The pre-crRNA contains nuclease guide sequences (spacers) interspaced by identical direct repeats (DRs). The Cas9 endonuclease can be engineered with a single 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. In comparison to other genome-editing technologies such as designer zinc fingers (ZFs), transcription activator-like effectors (TALEs) and homing meganucleases, the CRISPR/CAS9 system is a scalable, affordable and easy to engineer. Therefore, the anti-CRISPR/CAS9 antibody can be a useful tool for detecting CRISPR/CAS9 positively transfected cells, reveling DSB sites in the genome and in ChIP (Chromatin Immunoprecipitation) related assays.
Synonyms: CRISPR-associated protein-9 nuclease; Crisper; Crispr RNA
Storage: -20C
Application: Monoclonal Anti-CRISPR/CAS9 recognizes over-expression systems containing CAS9 construct. The antibody successfully recognizes mutant Cas9 variants, including nickase Cas9 and dead Cas9 (dCas9). The antibody may be used in various immunochemical techniques including Immunoblotting, Immunofluorescence and Immunoprecipitation. Monoclonal Anti-CRISPR/CAS9 does not cross react with FnCas9 from Francisella novicida bacteria and Cpf1 proteins from Acidaminococcus sp. (strain BV3L6) and Lachnospiraceae bacterium ND2006.
Biochem Physiol Actions: CAS9 plays a vital role in plasmid DNA interference. It is the only cas protein needed to deliver resistance against foreign DNA. CAS9 stimulates both RNA-guided genome editing and gene regulation in various organisms, but it can facilitate only one activity at a time within any given cell.
Other Notes: This product is for R&D use only, not for drug, household, or other uses. In order to obtain best results in different techniques and preparations we recommend determining optimal working concentration by titration test.