Huwe1 is a ubiquitin E3 ligase and tags various proteins for degradation via the ubiquitin-proteasome pathway. The aim of our study is to elucidate a role of Huwe1 in female infertility. Utilizing the Zp3-Cre/loxP system, we generated an oocyte-specific Huwe1 knockout (KO) mouse line. When Huwe1 heterozygous females were bred with wild type males, the number of litters was significantly reduced and, more importantly, the Huwe1 knockout allele was not passed on to the litters. In Huwe1 heterozygous females, oocytes matured and fertilized in vitro normally. However, only 50% of eggs isolated from Huwe1 heterozygous females reached the blastocyst stage in vitro. This suggests that Huwe1 knockout eggs died at an earlier stage. Interestingly, the oocyte-specific homozygous deletion of the Huwe1 gene caused complete female infertility. In vitro oocyte maturation assays further revealed that almost all of the knockout oocytes died or remained arrested at the germinal vesicle (GV) stage, whereas the majority of Huwe1 wild type oocytes matured to meiosis II. These results suggest that the loss of Huwe1 negatively affects oocyte maturation and early embryonic development, resulting in female infertility. Importantly, knockout of one of the well-known substrates of Huwe1, tumor suppressor p53, did not rescue any of these phenotypes in Huwe1 KO female mice. Therefore, it is suggested that female infertility in Huwe1 KO mice is secondary to the accumulation of a substrate of Huwe1 other than p53. Together, our results indicate that maternal Huwe1 is indispensable for oocyte maturation and early embryonic development.