Objective：To construct nerve injury-inducing protein 2（Ninj2） knockout mice and study its role during mouse ovarian development. Methods：We generated Ninj2 knockout mice by CRISPR/Cas9 technology，and detected the role of Ninj2 in mouse ovarian development by HE staining，immunofluorescence，in vitro fertilization（IVF） and lipopolysaccharide（LPS） exposure. Results：Ninj2 knockout mouse model was successfully constructed. Body weight and detection of related biochemical indicators showed that there was no significant difference between Ninj2 homozygous knockout mice and wild-type mice；HE staining showed that there was no significant difference in follicular development between Ninj2 knockout mice and wild-type controls at all levels in newborn and adult ovaries；IVF results showed that the early embryos of Ninj2 knockout mice developed normally；LPS had no effect on ovarian development in Ninj2 knockout mice. Conclusion：Depletion of Ninj2 does not affect the mouse ovarian development in normal physiological or inflammatory condition.