GenCRISPR™ ゲノム編集関連製品 » GenCRISPR™ ヌクレアーゼ-CRISPR遺伝子編集 » GenCrispr eSpCas9-N-NLS Nuclease
GenCrispr ESpCas9-N-NLS Nuclease

A 20 μl reaction in 1 × Cas9 Nuclease Reaction Buffer containing linearized plasmid, gRNA, and eSpCas9-N-NLS for 2 hour at 37 °C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.

GenCrispr eSpCas9-N-NLS Nuclease

GenCrispr eSpCas9 nuclease is a mutant form of Cas9 nuclease. Cas9 nuclease is an RNA-guided endonuclease that can catalyze cleavage of double stranded DNA. This kind of targeted nuclease is a powerful tool for genome editing with high precision. Cas9 protein forms a very stable ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component of the CRISPR/Cas9system. Compared with the wild type Cas9 nuclease, eSpCas9 reduces off-target effects by over 10-fold, while maintaining robust on-target genome editing efficiency. GenScript has developed a eSpCas9-N-NLS nuclease which contains a nuclear localization sequence (NLS) on the N-terminus of the protein to meet all the researchers’ requirements (e.g. in vitro cleavage assay, RNP complex transfection, and micro injection).
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Description

GenCrispr eSpCas9 nuclease is a mutant form of Cas9 nuclease. Cas9 nuclease is an RNA-guided endonuclease that can catalyze cleavage of double stranded DNA. This kind of targeted nuclease is a powerful tool for genome editing with high precision. Cas9 protein forms a very stable ribonucleoprotein (RNP) complex with the guide RNA (gRNA) component of the CRISPR/Cas9system. Compared with the wild type Cas9 nuclease, eSpCas9 reduces off-target effects by over 10-fold, while maintaining robust on-target genome editing efficiency. GenScript has developed a eSpCas9-N-NLS nuclease which contains a nuclear localization sequence (NLS) on the N-terminus of the protein to meet all the researchers’ requirements (e.g. in vitro cleavage assay, RNP complex transfection, and micro injection). _x000D_

_x000D_ Product Source: GenCrispr eSpCas9-N-NLS is produced by expression in an E. coli strain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes with a N terminal nuclear localization signal (NLS)._x000D_

_x000D_ Key Features _x000D_
_x000D_ -DNA-free: no external DNA added to system_x000D_
_x000D_ -High cleavage efficiency: NLS ensures the entry of Cas9 protein into nuclei_x000D_
_x000D_ -Low off target: transient expression of Cas9 nuclease_x000D_
_x000D_ -Time-saving: no need for transcription and translation_x000D_

_x000D_ Applications _x000D_
_x000D_ -Screening for highly efficient and specific targeting gRNAs by in vitro DNA cleavage. _x000D_
_x000D_ -In vivo gene editing when combined with a specific gRNA by electroporation or injection.
Note

This is a basic protocol. The reagent concentrations, conditions, and parameters may need to be optimized.
1000 nM is equal to 160 ng/μL.
For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.

Properties
Storage & Stability GenCrispr eSpCas9-N-NLS nuclease is supplied with 1X storage buffer (10 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol PH 7.4, at 25°C) and recommended to be stored at -20°C. Guaranteed stable for 18 months when properly stored.

Examples
  • GenCrispr ESpCas9-N-NLS Nuclease
    • GenCrispr ESpCas9-N-NLS Nuclease

    A 20 μl reaction in 1 × Cas9 Nuclease Reaction Buffer containing linearized plasmid, gRNA, and eSpCas9-N-NLS for 2 hour at 37 °C results in a digestion efficiency of linearized plasmid higher than 90%, as determined by agarose gel electrophoresis.