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| Z03699 | |
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The GenCRISPR™ SaCas9 2NLS Nuclease can be formed with the guide RNA into a ribonucleoprotien (RNP) complex. The use of an RNP complex to perform gene editing has been shown to reduce the challenges encountered with other CRISPR gene editing techniques such as viral and plasmid delivery. Challenges include off-target effects, cell viability and transcription/translational challenges. The SaCas9 recognizes an NNGRRT protospacer adjacent motif (PAM) and cleaves target DNA at high efficiency with a variety of guide RNA (gRNA) spacer lengths. GenCRISPR™ SaCas9 2NLS Nuclease is a tag free nuclease produced by expression in an E. coli strain carrying a plasmid encoding the Cas9 gene from Staphylococcus aureus with a nuclear localization signal at both N-terminal and C-terminal.The small size of the nuclease facilitates enhanced in vivo delivery for genome editing in various organisms. |
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.
| Key Features |
High knockout
efficiencies: Consistent high performance in in-vitro plasmid cleavage test. Tag-free: No extra tag amino acid. DNA-free: No external DNA added to the system. |
|
gRNA-dependent double-stranded DNA cleavage |
| Expression System |
Recombinant
SaCas9 with an NLS at both N-terminal and C-terminal expressed by E.coli |
| Species |
Staphylococcus aureus |
| Tag |
Tag-free |
| Molecular Weight |
~130
kDa |
| Concentration |
4 mg/ml |
| Active temperature |
This
SaCas9 is active at 37°C. |
| Formulation |
Supplied as a solution of 20 mM Tris, 300 mM NaCl, 0.1
mM TCEP, 50% glycerol, pH 7.5 |
| Storage & Stability | This product remains stable up to 12 months at -20°C. Avoid repeated freeze-thaw cycles |
| Appearance |
Clear,
colorless liquid |
| Purity |
≥ 90%
as analyzed by SDS-PAGE |
| Concentration |
4
mg/ml±10% |
| Bioactivity |
≥ 90% |
| Residual DNase |
Non-specific
DNase activity |
| Residual RNase |
Non-specific
RNase activity |
| Endotoxin Level |
≤
100 EU/mg as analyzed by gel clotting method |
A 20 μl reaction in 1 × SaCas9 Nuclease Reaction Buffer containing linearized plasmid, gRNA, and SaCas9 for 2 hours at 37 °C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis. »
