GenCRISPR™ ゲノム編集関連製品 » GenCRISPR™ ヌクレアーゼ-CRISPR遺伝子編集 » GenCRISPR™ Ultra NLS-Cas9-Research
GenCRISPR™ Ultra NLS-Cas9-Research

A 20 μl reaction in 1 × Cas9 Nuclease Reaction Buffer containing 160 ng linearized plasmid, 100 ng gRNA, and 50 ng GenCRISPR™ Ultra Cas9 for 2 hours at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.

GenCRISPR™ Ultra NLS-Cas9-Research

GenCRISPR™ Ultra Cas9 is produced by expression in an E. coli strain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes with an N-terminal nuclear localization signal (NLS).
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Description

The_x000D_ GenCRISPR™ Ultra Cas9 product line provides customers with a selection of_x000D_ research use, GMP preclinical and GMP compliant Cas9 nucleases. The Cas9_x000D_ protein can be formed with the guide RNA into a ribonucleoprotien (RNP)_x000D_ complex. The use of an RNP complex to perform gene editing has been shown to_x000D_ 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 trascprition/translational challenges.

_x000D_ _x000D_ _x000D_ _x000D_ GenCRISPR™ Ultra Cas9 is_x000D_ produced by expression in an E. coli_x000D_ strain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes with an N-terminal nuclear localization_x000D_ signal (NLS).
Note

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

Overview
Key Features High knockout efficiencies: Consistent high editing efficiency in in-vitro and in-vivo.
Tag-free: Amino acid is free from additional tagging amino acid.
Animal-free: Plant-based culture media, enzyme-free chromatographic purification.
DNA-free: No external DNA added to the system.

Applications
gRNA-dependent double-stranded DNA cleavage

Properties
Expression System Recombinant Cas9 with an N-terminal NLS expressed by E.coli
Species S. pyogenes
Tag Tag-free
Molecular Weight ~160 kDa
Concentration 10 mg/mL
Active temperature This Cas9 is active at 37 ℃.
Formulation Supplied as a solution of 25 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 50% Glycerol pH 8.0 at 25°C. 
Storage & Stability This product remains stable up to 12 months at -20°C. Avoid repeated freeze-thaw cycles.

Quality Control Specifications
Appearance Clear, colorless liquid
Purity ≥ 95% as analyzed by SDS-PAGE
≥ 90% as analyzed by SEC-HPLC
Concentration by A280 10 mg/ml ± 1 mg/ml
Bioactivity (in vitro) ≥ 90%
Residual DNase Non-specific DNase activity
Residual RNase Non-specific RNase activity
Endotoxin Level ≤ 10 EU/mg as analyzed by gel clotting method

Examples
  • GenCRISPR™ Ultra NLS-Cas9-Research
    • GenCRISPR™ Ultra NLS-Cas9-Research

    A 20 μl reaction in 1 × Cas9 Nuclease Reaction Buffer containing 160 ng linearized plasmid, 100 ng gRNA, and 50 ng GenCRISPR™ Ultra Cas9 for 2 hours at 37°C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.