Abstract
Deriving from the adaptive immune system of prokaryotes, the emerging gene-editing technology CRISPR/Cas9, has become the most popular and powerful tool for the manipulation of genes at the molecular level. It has the advantages of easy construction, high efficiency along with good specificity. CRISPR/Cas9 system has been extensively applied to model constructions, clinical treatment and mechanisms of tumorigenesis. As a number of strategies for CRISPR/Cas9 system have been developed, this review systematically summarizes the commonly applied methods along with discussions on their advantages and limitations and different scenarios for specific applications. The basic delivery forms include plasmid DNA with sgRNA, mRNA with sgRNA and a complex of Cas9 protein and sgRNA, ribonucleoprotein (RNP). For the methods of delivery into cells, there are viral vectors and non-viral vectors, which further consist of physical and chemical methods. While the efficiency and rate of successful transfection vary in different strategies, they commonly suffer from off-target effects, mutagenesis, cytotoxicity and immunogenicity to different extents.
Publisher
Darcy & Roy Press Co. Ltd.
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