Method for constructing zebrafish glycogen storage disease gys1 and gys2 gene mutant

The invention discloses a method for constructing a zebrafish glycogen storage disease gys1 and gys2 gene mutant. The method includes the following steps of selecting a fourth exon sequence of a zebrafish gys1 gene sequence to design a gys1 gene knockout target, selecting a third exon sequence of a gys2 gene sequence to design a gys2 gene knockout target, designing an amplification primer according to the above target sequences, conducting PCR amplification with pUC19-scaffold plasmid as a template, conducting purification and in vitro transcription to obtain the gys1 and gys2 gene knockout target gRNA, and importing the gRNA and a Cas9 protein into a zebrafish fertilized egg to obtain the stable genetic zebrafish gys1 and gys2 gene homozygous mutant. Zebrafish has large egg production, short breeding cycle and transparent embryo bodies, and the method is favorable for further study of the pathogenesis and detailed mechanism of human glycogen storage diseases. 本发明公开了斑马鱼糖原贮积症gys1和gys2基因突变体的构建方法，包括以下步骤：选择斑马鱼gys