Application and Comparison of CRISPR-Cas9 System and CRISPR-Cpf1 System in Multigenome Editing
GUO Ting, AN Xinmin
Gene editing technology is a kind of technology that introduces nucleic acid sequence changes into gene target, which has been widely used in biology, basic medicine and other fields. With the deepening of the research on CRISPR system, the multiple gene editing technology using Cas9 protein has developed rapidly. Scientists have developed a variety of multiple gRNAs vector construction strategies relying on Cas9 protein, which has realized multiple gene editing in various species. CRISPR-Cpf1 system is a new tool of gene editing technology, which has greatly enriched the CRISPR/Cas system library. It not only further expands the selection range of gene editing target sites, and causes less off-target effects, but also its molecular mechanism of action different from Cas9 protein confers it natural advantage in multiple gene editing, which has attracted extensive attention. In this paper, we mainly introduced four construction strategies of multiple gene editing based on CRISPR Cas9 system, including Golden Gate Assembly, Multiplexed Lentiviral Expression Cassettes, Polycistronic-tRNA-Grna Cassettes, Csy4-cleavable Cassettes, and CRISPR-Cpf1 multi-gene editing technology. Meanwhile, we compare the characteristics of CRISPR-Cas9 system and the CRISPR - Cpf1 system, and expect to provide reference for the application of multiple gene editing technology in the field of biological research.
CN
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