GenCRISPR™ ゲノム編集関連製品 » GenCRISPR™ ヌクレアーゼ-CRISPR遺伝子編集 » GenCRISPR™ SaCas9 2NLS Nuclease
GenCRISPR™ SaCas9 2NLS Nuclease

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.

GenCRISPR™ SaCas9 2NLS Nuclease

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.
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Description

The GenCRISPR™_x000D_ SaCas9 2NLS Nuclease can be formed with the guide RNA into a ribonucleoprotien_x000D_ (RNP) complex. The use of an RNP complex to perform gene editing has been shown_x000D_ to reduce the challenges encountered with other CRISPR gene editing techniques_x000D_ such as viral and plasmid delivery. Challenges include off-target effects, cell_x000D_ viability and transcription/translational challenges. The SaCas9 recognizes an_x000D_ NNGRRT protospacer adjacent motif (PAM) and cleaves target DNA at high_x000D_ efficiency with a variety of guide RNA (gRNA) spacer lengths.

_x000D_ _x000D_ _x000D_ _x000D_ GenCRISPR™ SaCas9 2NLS_x000D_ Nuclease is a tag free nuclease produced by expression in an E. coli strain carrying a plasmid_x000D_ encoding the Cas9 gene from Staphylococcus_x000D_ aureus with a nuclear localization signal at both N-terminal and C-terminal.The_x000D_ small size of the nuclease facilitates enhanced in vivo delivery for genome_x000D_ editing in various organisms.
Note

For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.

Applications
gRNA-dependent double-stranded DNA cleavage

Properties
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
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.

Quality Control Specifications
Appearance Clear, colorless liquid
Purity ≥ 90% as analyzed by SDS-PAGE
Concentration by A280 4 mg/ml±10%
Bioactivity (in vitro) ≥ 90%
Residual DNase Non-specific DNase activity
Residual RNase Non-specific RNase activity
Endotoxin Level ≤ 100 EU/mg as analyzed by gel clotting method

Examples
  • GenCRISPR™ SaCas9 2NLS Nuclease
    • GenCRISPR™ SaCas9 2NLS Nuclease

    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.