Objective: To study the effect and mechanism of Tip60 regulating DNA on mouse embryo development, and to provide theoretical basis for the study of embryo development. Methods: mice embryos were randomly divided into two groups: control group and experimental group. The embryos of experimental group were given Tip60 inhibitor, the mice of control group were given the same dose of normal saline. The level of Tip60 in the two groups of mice embryos, the effect of Tip60 on p53-p21 pathway, the expression of DNA repair factor 53BP1 protein, the content of active oxygen, the effect of autophagy and apoptosis, and the development of embryos were analyzed. Results: the level of Tip60 in the experimental group was significantly lower than that in the control group; the p53-p21 pathway in the experimental group was activated, the DNA damage of the experimental group was greater, the expression of DNA repair factor was lower, the content of ROS in the experimental group was significantly higher than that in the control group; the autophagy and apoptosis of the experimental group were enhanced; the capsule of the control group was enhanced. The embryogenesis rate of the experimental group was (65.13 ± 4.85)％, and that of the experimental group was (29.36 ± 1.75)％, which was significantly lower than that of the control group. Conclusion: Tip60 can regulate DNA damage and repair by mediating p53-p21 pathway in early mouse embryo development. Inhibition of Tip60 can increase the active oxygen, induce autophagy and over expression of apoptosis in mouse embryo cells, and inhibit the development of embryo to a certain extent. It has important guiding significance to measure the level of Tip60 during embryo culture.