The Development of Multi-Gene Editing Technology and Its Application in Agricultural Biological Germplasm Innovation
CRISPR gene editing technology can accurately and efficiently change the DNA source code and has been widely used in animal and plant breeding in recent years.In practice,the de-mand for multi-trait parallel improvement of agricultural organisms is increasing.The change of a single gene or locus alone cannot meet the above requirements.Therefore,it is desirable to es-tablish a multi-gene simultaneous editing system to modify multiple genes.Simultaneous expres-sion of multiple sgRNAs is critical to simultaneous editing of multiple genes.The commonly multi-sgRNA expression strategies include multiple single-cistron sgRNA parallel expression and multi-cistron sgRNA tandem expression.The frequently-used tandem expression tools include nuclease Csy4,tRNA system and self-cleaving ribozyme.This review analyzes and summarizes the advantages and disadvantages of the above multi-gene editing technology,discusses the future direction,and stresses its significance and application prospects.
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