Mobile transposon-like element, clone MTi7: Finding its role(s) by RNA interference. |
Chang Eun Park, Mi Ra Shin, Eun Hyun Jeon, Sung Won Cho, Sook Hwan Lee, Kyungjin Kim, Nam Hyung Kim, Kyung Ah Lee |
1Infertility Medical Center, CHA General Hospital, Korea. 2Graduate School of Life Science and Biotechnology, Pochon CHA University, College of Medicine, Korea. 3Department of Animal Science, Chungbuk National University, Korea. 4School of Biological Sciences, Seoul National University, Korea. |
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Abstract |
OBJECTIVES: The present study was conducted to evaluate the mobile transposon-like element, clone MTi7 (MTi7) expression in the mouse ovary and to determine its role(s) in the mouse oocytes by RNA interference (RNAi). METHODS: MTi7 mRNA expression was localized by in situ hybridization in day5 and adult ovaries. Double stranded RNA (dsRNA) was prepared for c-mos, a gene with known function as control, and the MTi7. Each dsRNA was microinjected into the germinal vesicle (GV) stage oocytes then oocyte maturation and intracellular changes were evaluated. RESULTS: In situ hybridization analysis revealed that MTi7 mRNA localized to the oocyte cytoplasm from primordial to preovulatory follicles. After dsRNA injection, we found 43-54% GV arrest of microinjected GV oocytes with 68%-90% decrease in targeted c-mos or MTi7 mRNA. CONCLUSIONS: This is the first report of the oocyte-specific expression of the MTi7 mRNA. From results of RNAi for MTi7, we concluded that the MTi7 is involved in the germinal vesicle breakdown in GV oocytes, and MTi7 may be implicated with c-mos for its function. We report here that RNAi provides an outstanding approach to study the function of a gene with unknown functions. |
Key Words:
MTi7; Mouse; Oocyte Maturation; RNA Interference |
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