Cas Enzyme Expression
About CRISPR-Ca9:
Cas9 (CRISPR-associated protein 9) is an endonuclease capable of cutting double-stranded DNA, enabling precise alterations to a cell’s genome. The CRISPR-Cas9 gene-editing system uses a guide RNA (gRNA) to direct the Cas9 enzyme to a specific target site within the genomic DNA. Once bound, Cas9 introduces a double-strand break (DSB) at the target location.
They are many Cas9 enzymes used in CRISPR. SpCas9, SaCas9 and Cas12a (Cpf1) are most frequently used and verified.
(1) SpCas9 (from Streptococcus pyogenes ) is the most frequently used cas endonuclease in CRISPR. It PAM sequence is: 5’-NGG-3’. Ca9 protein can be delivered as the standalone expression, or delivered with a target gRNA in a one-vector system.
(2) SaCas9 (from Staphylococcus aureus) is a smaller Cas9 ortholog (~3.2 kb) than SpCas9 (4.2kb). The SaCas9 use the PAM sequence as: 5′-NNGRRT-3‘ (R = A or G). The SaCas9 is often has less off-target effects than SpCas9.
(3) Cas12a (Cpf1) (from Acidaminococcus sp) is a Class 2, Type V CRISPR-associated endonuclease used for genome editing. Its compact size and distinct PAM [ TTTV (V = A/C/G)] and cleavage features make Cas12a a powerful alternative to Cas9 for precise and flexible gene editing across a variety of organisms.
GenTarget offers standalone SpCas9, SaCas9, AsCas12a Expression Lentivirus, driven by different promoters with a variety of antibiotic selection markers (see the core expression lentivector scheme below).
Cas9 Lentivirus Product Features:
- High Cas9 expression Level by enhanced promoters to fit different cell types;
- High titer lentivirus provides more efficient Cas9 delivery in almost all cell types including primary cells and non-dividing cells;
- Flexible selection with different antibiotic and fluorescent markers;
- Best nuclear penetrating with enhanced NLS leader: the Cas9 is expressed with an optimized, proprietary Nuclear Localization Signal (NLS), providing the efficient cas9 delivery into the nuclear region where the gene-editing occurs.
- Easy to use: you can simply synthesize the gRNA (or use gRNA lentivirus) and used it together with the Cas9 lentivirus for targeted gene editing.
- The standalone Cas9 lentivirus allows generating CRISPR-ready animal models or stable cell lines in your desired cell types or animals that are used as a model for any gene editing.
Please see details in SpCas9 Product Manual, SaCas9 Product Manual, or AsCas12a Product Manual.
Gentarget also provides the SpCas9 Expression Adenovirus, SaCas9 Expression AAV virus, and Cand the Integration-Deficient Lentivirus for Cas9 expression.