@misc{oai:repo.qst.go.jp:00069106, author = {塚本, 智史 and 今市, 寿史 and 太田, 有紀 and 鬼頭, 靖司 and 南, 直治郎 and その他 and 太田 有紀 and 鬼頭 靖司 and 南 直治郎}, month = {Jul}, note = {Gene products stored in oocytes play important roles after the completion of meiosis and fertilization. These genes are called maternal effect gene and shown to be important in early embryonic development in many species. We previously identified Oog1 that specifically expressed in oocytes at all stages and preimplantation embryo by the late two-cell stage and showed that Oog1 protein localized in nuclei at the late one-cell and early two-cell stages, the time of the occurrence of zygotic genome activation (ZGA) in mice. We also reported that Oog1 interacts with Ras effectors (e.g., RalGDS and Rasa4) and also binds to Ras in a GTP dependent manner. These results suggested that Oog1 is one of the Ras mediated signaling proteins. However, the biological function of Oog1 in oogenesis and/or early embryogenesis is still unclear. In the present study, we attempted to carry out Oog1 knockdown experiments using RNAi. We first constructed a long-hairpin dsRNA (about 500 bp) that target Oog1 (Oog1-hairpin dsRNA). To check if the constructed dsRNA was capable of inhibiting maternal Oog1 mRNA specifically, GV-oocytes were microinjected with in vitro transcribed dsRNA fused with EGFP. Twenty hours after microinjection, RT-PCR analysis was performed in EGFP-positive oocytes showing that Oog1 mRNA was dramatically reduced. The dsRNA did not affect the amount of Oog2, 3 and 4, which are highly homologous to Oog1 and expressed in the oocyte. These results suggest that the Oog1-hairpin dsRNA effectively and specifically reduces Oog1 mRNA in oocytes. Because Oog1 mRNA and its protein start to express at the beginning of oogenesis, we employed a recently developed transgenic RNAi approach using an oocyte-specific Zp3 promoter to reduce Oog1 mRNA in growing oocytes. From the thirty founder mice, five transgenic female and four transgenic male were obtained, all of which are apparently normal. To obtain F1 transgenic progeny, transgenic female and male founder were mated with wild-type male and female mice, respectively. We are currently investigating the phenotype (including fertility) in the transgenic female and F1 female progeny., The 40th annual Meeting, Society for the Study of Reproduction}, title = {RNAiによる卵子特異的遺伝子Oog1の効率的な抑制}, year = {2007} }