Description
Pre-packaged rAAV in Serotype-8, constitutively express the human codon-optimized Staphylococcus aureus Cas9 (SaCas9) under a modified CMV Promoter, with N-terminal NLS (Nuclear Localization Signal) for nuclear penetrating, and C-terminal 6His tag. The expressed SaCas9 can be detected by anti-C-terminal 6His tag antibody. The following scheme showed the AAV vector’s core structure. See Product Manual for details.
Streptococcus pyogenes Cas9 (SpCas9) is the most commonly used CRISPR endonuclease, but its large coding sequence (~4.2 kb) exceeds or near the packaging capacity of AAV virus, which has a maximum of ~4.7–5.0 kb including all regulatory elements.
To overcome this limitation, Staphylococcus aureus Cas9 (SaCas9) is used instead. SaCas9 is a smaller Cas9 ortholog (~3.2 kb), allowing it to fit within AAV vectors. Both are CRISPR-associated endonucleases used for genome editing. However, their PAM sequence is different. The SaCas9 use the PAM sequence as: 5′-NNGRRT-3‘ (R = A or G) whereas the SpCas9 as: 5’-NGG-3’. The SaCas9 is often has less off-target effects than SpCas9. see also Gentarget’s SpCas9 expression Lentivirus products.
AAV Serotype-8 has a higher preference for transducing Liver (hepatocytes), Heart, Skeletal Muscle, CNS, Kidney, pancreas. To determine the serotype that best suits your desired cell types, you can use the AAV-GFP Serotype Screening Kit.
Titer: ~ 1×10^12 GC/mL
Size: 100ul/vial, in stock, ready to ship in overnight Dry-ice package.
Storage Buffer: PBS
CAT#: A11.8
