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GenCrispr NLS-Cas9-EGFP Nuclease

GenCrispr NLS-Cas9-EGFP Nuclease

In vitro DNA cleavage assay with GenCrispr NLS-Cas9-EGFP nuclease
Reactions were set up according to recommended conditions, and cleavage products were resolved on a 1% agarose gel. Input DNA is EcoR V-linearized pUC57 plasmid DNA. Lane 1, DNA + gRNA; lane 2, marker; lane 3 and 4, DNA + gRNA + NLS-Cas9-EGFP 100 ng (lane 3) or 50 ng (lane 4).

GenCrispr NLS-Cas9-EGFP Nuclease

Left: Cell transfection assay. 12 h after electrophoresis, cells were observed under bright or fluorescence microscope.
Right: in vivo gene editing efficiency assay by T7E1. Lane 1, marker; lane 2, negative control; lane 3, gRNA + NLS-Cas9-EGFP; lane 4, gRNA + NLS-Cas9.



Description
GenCrispr NLS-Cas9-EGFP is a fusion protein developed by GenScript. It contains a nuclear localization sequence (NLS) on its N terminal and EGFP on the C terminal. 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 or translation compared with mRNA or plasmid systems. Additionally, the Cas9 RNP complex is rapidly cleared from the cells minimizing the chance of off-target cleavage when compared to other systems (Kim, et al. 2014). The EGFP can be taken as a reporter for tracking or sorting transfected cells, which creates the possibility of enriching cell populations for desired genome edits via fluorescence activated cell sorting (FACS). It significantly reduces the labor and cost associated with single cell cloning and genotyping in genome editing applications.

Product Source: GenCrispr NLS-Cas9-EGFP is produced by expression from an E. coli strain.
KEY FEATURES
  • DNA-free: no external DNA added to system
  • High cleavage efficiency: NLS ensures the efficient entry of Cas9 protein into nuclei
  • Low off target: transient expression of Cas9 nuclease
  • Time-saving: no need for transcription and translation
  • Reduced-labor: enrich cell populations for desired genome edits via EGFP-based fluorescence activated cell sorting (FACS).
Utilities of Product
  • Screening for highly efficient and specific targeting gRNAs by in vitro DNA cleavage.
  • In vivo gene editing when combined with a specific gRNA by electroporation or injection.
  • Enrich cell populations for desired genome edits via EGFP-based fluorescence activated cell sorting (FACS).

Storage GenCrispr NLS-Cas9-EGFP 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.

GenCrispr NLS-Cas9-EGFP Nuclease
GenCrispr NLS-Cas9-EGFP Nuclease

In vitro DNA cleavage assay with GenCrispr NLS-Cas9-EGFP nuclease
Reactions were set up according to recommended conditions, and cleavage products were resolved on a 1% agarose gel. Input DNA is EcoR V-linearized pUC57 plasmid DNA. Lane 1, DNA + gRNA; lane 2, marker; lane 3 and 4, DNA + gRNA + NLS-Cas9-EGFP 100 ng (lane 3) or 50 ng (lane 4).

GenCrispr NLS-Cas9-EGFP Nuclease
GenCrispr NLS-Cas9-EGFP Nuclease

Left: Cell transfection assay. 12 h after electrophoresis, cells were observed under bright or fluorescence microscope.
Right: in vivo gene editing efficiency assay by T7E1. Lane 1, marker; lane 2, negative control; lane 3, gRNA + NLS-Cas9-EGFP; lane 4, gRNA + NLS-Cas9.




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Cat. No. Z03393
Size
Price
50 µg (1mg/ml) $179.00
100 µg (3mg/ml) $229.00
24 hour
 
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