GenCrispr NLS-Cas9-EGFP Nuclease

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 and 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 used as a reporter for tracking or sorting transfected cells, which enables 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.