In vitro DNA cleavage assay with Z03469.
Reactions were set up according to recommended conditions, and cleavage products were resolved on a 1.5% agarose gel. Input DNA is linearized pUC57 plasmid DNA. The cleavage efficiency of Z03469 is comparable to competitors.
Lane 1, marker;
Lane 2-3, DNA + gRNA + IDT;
Lane 4-5, DNA + gRNA + Merck;
Lane 6-7, DNA + gRNA + Z03469;
Lane 8-9, negative control.

In vivo gene editing efficiency assay of Z03469:
Cas9 and gRNA were transfected into THP1 cells by electroporation. Cells were lysed after 48h and analyzed by T7E1 assay. The editing efficiency of Z03469 is higher than competitors.
Lane 1, gRNA + Z03469;
Lane 2, gRNA + Merck;
Lane 3, gRNA + IDT;
Lane 4, gRNA + Thermo;
Lane 5, negative control;
Lane 6, marker.

In vivo gene editing efficiency assay of Z03469:
Cas9 and gRNA were transfected into 293T cells by liposome. Cells were lysed after 48h and analyzed by T7E1 assay. The editing efficiency of Z03469 is comparable to competitors.
Lane 1, marker;
Lane 2, gRNA + Z03469;
Lane 3, gRNA + Merck;
Lane 4, gRNA + IDT;
Lane 5, negative control.

GenCrispr NLS-Cas9-NLS 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/Cas9 system. The Cas9 RNP complex can localize to the nucleus immediately upon entering the cell with the addition of a nuclear localization signal (NLS). There is no requirement for transcription and translation compared with mRNA or plasmid systems. Additionally, the Cas9 RNP complex is rapidly cleared from the cell minimizing the chance of off-target cleavage when compared to other systems (Kim, et al. 2014). This DNA-free system avoids the risk of inserting foreign DNA into the genome, which can be quite useful for gene editing-based disease therapy. GenScript has developed a NLS-Cas9-NLS nuclease which contains a nuclear localization sequence (NLS) on both ends of the protein to meet all the researchers’ requirements (e.g. in vitro cleavage assay, RNP complex transfection, and micro injection).

Cat. No.	Z03469
Size	100 μg (10mg/ml) 500 μg (10mg/ml) 1 mg (10mg/ml) Customized Size
Quantity
Price	¥34,075.00


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Description

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/Cas9 system. The Cas9 RNP complex can localize to the nucleus immediately upon entering the cell with the addition of a nuclear localization signal (NLS). There is no requirement for transcription and translation compared with mRNA or plasmid systems. Additionally, the Cas9 RNP complex is rapidly cleared from the cell minimizing the chance of off-target cleavage when compared to other systems (Kim, et al. 2014). This DNA-free system avoids the risk of inserting foreign DNA into the genome, which can be quite useful for gene editing-based disease therapy. GenScript has developed a NLS-Cas9-NLS nuclease which contains a nuclear localization sequence (NLS) on both ends 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 NLS-Cas9-NLS is produced by expression in an E. coli strain carrying a plasmid encoding the Cas9 gene from Streptococcus pyogenes with a nuclear localization signal (NLS) on both ends. _x000D_

_x000D_ Key Features _x000D_
_x000D_ -DNA-free: no external DNA added to system_x000D_
_x000D_ -High cleavage efficiency: Double NLS ensures the efficient 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.

Properties

Storage & Stability

GenCrispr NLS-Cas9-NLS Nuclease is supplied with 1X storage buffer (10 mM Tris, 300 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol pH7.4, at 25°C) and recommended to be stored at -20°C. Guaranteed stable for 18 months when properly stored.

Examples