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GenCRISPR™ Cas9 v1.1

*This product has been discontinued!*
GenCRISPR™ Cas9 v1.1 is a tag free nuclease produced by expression in an E. coli strain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes with a biparticle nucleus localization signal (BPNLS) at both N-terminal and C-terminal. It has been reported that BPNLS is able to improve the gene editing efficiency.
Z03701
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Description

The GenCRISPR™ Cas9 v1.1 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.

GenCRISPR™ Cas9 v1.1 is a tag free nuclease produced by expression in an E. coli strain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes with a biparticle nucleus localization signal (BPNLS) at both N-terminal and C-terminal. It has been reported that BPNLS is able to improve the gene editing efficiency.

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: No extra tag amino acid. 
DNA-free: No external DNA added to the system.
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Applications
gRNA-dependent double-stranded DNA cleavage
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Properties
Source Recombinant Cas9 with a BPNLS at both N-terminal and C-terminal expressed by E.coli 
Species Streptococcus pyogenes
Tag Tag-free
Molecular Weight ~160 kDa
Concentration 4 mg/ml
Active temperature This Cas9 is active at 37 °C.
Formulation Supplied as a solution of 25 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 50% glycerol, pH 8.0.
Storage & Stability This product remains stable for up to 12 months at -20 °C. Avoid repeated freeze-thaw cycles.
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Quality Control Specifications
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
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Background
References 1.      Liu, Xinyi, et al. "Improving editing efficiency for the sequences with NGH PAM using xCas9-derived base editors." Molecular Therapy-Nucleic Acids 17 (2019): 626-635. 
2.      Pollard, Victoria W., et al. "A novel receptor-mediated nuclear protein import pathway." Cell 86.6 (1996): 985-994. 
3.      Koblan, Luke W., et al. "Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction." Nature biotechnology 36.9 (2018): 843-846.

--> 1.      Liu, Xinyi, et al. "Improving editing efficiency for the sequences with NGH PAM using xCas9-derived base editors." Molecular Therapy-Nucleic Acids 17 (2019): 626-635. 
2.      Pollard, Victoria W., et al. "A novel receptor-mediated nuclear protein import pathway." Cell 86.6 (1996): 985-994. 
3.      Koblan, Luke W., et al. "Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction." Nature biotechnology 36.9 (2018): 843-846.
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