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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