Objective:This study aims to observe the effect of microRNA-1 (miR-1) on the development of neural crest cells and craniofacial region in the knockout zebrafish model. Methods: miR-1 of zebrafish was knocked out by CRISPR / Cas9 gene knock-out system. In situ hybridization was used to detect the expression of differentiation related genes in neural crest cells, and qRT-PCR was used to detect the expression of apoptosis related genes. Results: In the knockout group, the mandible was atrophic and the pigment cells appeared delayed. In situ hybridization showed that the expression of tfap2a,dlx3b, ngn1 and snailb decreased 24 hours after fertilization. qRT-PCR results showed that miR-1 affected the expression of apoptosis related genes. Conclusion: miR-1 is involved in the regulation of neural crest cells and affects craniofacial development, which may play a role by regulating apoptosis related pathways.