SaCas9 Nuclease
Product Introduction
SaCas9 nuclease, derived from Staphylococcus aureus, is one of the most widely used small-sized Cas9 orthologs and holds particular value in the field of gene therapy. This protein consists of 1053 amino acids with a molecular weight of approximately 120 kDa, which is about 25% smaller than SpCas9 (1368 amino acids).
SaCas9 recognizes the PAM sequence 5'-NNGRRT-3' (where R represents A or G), which is longer and more complex than the NGG PAM recognized by SpCas9. This longer recognition sequence theoretically enhances targeting specificity and reduces off-target risks, as fully matched sites occur less frequently in the genome. Studies have shown that SaCas9 exhibits robust editing activity in mammalian cells, with cleavage efficiency comparable to or even better than SpCas9.
Product Specifications
Parameter | Specification |
Source | Recombinant expression in E. coli |
Molecular Weight | ~120 kDa |
Concentration | 20 μM |
PAM Sequence | 5'-NNGRRT-3' (R = A/G) |
Cleavage Site | 3 bp upstream of PAM |
Cleavage Product | Blunt-ended DSB |
Purity | ≥95% (SDS-PAGE) |
Endotoxin | <1 EU/μg |
Storage Buffer | 50 mM Tris-HCl, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol |
10× Reaction Buffer | 50 mM Tris-HCl, 100 mM NaCl, 10 mM MgCl₂, 100 μg/ml BSA, pH 7.9 |
Storage Conditions | Long-term storage at -80°C; short-term storage at -20°C |
Product Specifications
Specifications | Catalog Number | Concentration | Volume |
100 pmol | GR101401 | 20 μM | 5 μL |
500 pmol | GR101402 | 20 μM | 25 μL |
2500 pmol | GR101403 | 20 μM | 125 μL |
Application Scenarios
AAV-Mediated In Vivo Gene Editing: Small size suitable for single AAV vector delivery.
Gene Therapy: AAV-based gene therapy for ophthalmic diseases, neurodegenerative disorders, etc.
Tissue-Specific Editing: Achieving tissue targeting by combining with AAV serotypes.
Dual Nickase Strategy: Paired use of SaCas9 D10A/N580A nickases.
Orthogonal Editing: Combined use with SpCas9 for dual gene editing.
References
Ran FA, et al. (2015). In vivo genome editing using Staphylococcus aureus Cas9. Nature. 520(7546):186-191.
Nishimasu H, et al. (2015). Crystal structure of Staphylococcus aureus Cas9. Cell. 162(5):1113-1126.
Friedland AE, et al. (2015). Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications. Genome Biol. 16:257.
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