Objective To study the detailed process and mechanism of glycogen storage disease (GSD) type 0 using a zebrafish disease model established by CRISPR/ Cas9 gene editing technology. Methods Target sites were determined for gys1 and gys2. After gene editing, homozygous mutants were screened. The expression levels of gys1 and gys2 were quantified with qPCR at early developmental and adult stages. Glycogen accumulation was assessed by periodic acid-Schiff staining. Results Homozygous F₂ gys1 and gys2 genes with two effective mutation types were successfully obtained. Homozygous gys1^{- / -} F₃ had a delay in early embryonic development and died within 48 hpf, whereas homozygous gys2^{- / -} developed normally. Gene expression analyses of gys1 and gys2 in wildtype zebrafish showed that their expression levels decreased first and then increased during early developmental stages (0~ 125 hpf) with the highest expression at the fertilized egg stage. At the adult stage, gys1 was highly expressed in heart and muscle tissues, while gys2 was highly expressed in the liver. Glycogen staining showed no significant glycogen accumulation in the heart or muscle of gys1^{- / -} or in the liver of gys2^{- / -} compared with the corresponding wildtype tissues. Conclusions The phenotypes of gys1 and gys2 knockout zebrafish models were similar to those of mouse models, but there were some differences. In the Gys1 knockout mouse model, most F₂ homozygotes die. Among gys1 knockout zebrafish, gys1^{- / -} homozygous F₂ developed normally, whereas F₃ generated by self-crossing did not survive. The model of gys1 and gys2 knockout in zebrafish provides materials for further study on the pathogenesis of human GSD type 0.