RNAi-knockdown analysis of a germ-cell-specific antigen, TEX101, in mouse testis
| Project/Area Number |
18591786
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | Nippon Medical School |
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Principal Investigator |
ISHIKAWA Tomoko Nippon Medical School, Faculty of Medicine, Assistant Professor (70212850)
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| Co-Investigator(Kenkyū-buntansha) |
TAKIZAWA Toshihiro Nippon Medical School, Graduate School of Medicine, Professor (90271220)
水口 義昭 日本医科大学, 医学部, 助手 (70409217)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,790,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | mouse testis / TEX101 / GPI / RNAi / reproductive biology / 精巣 / 精子 |
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| Research Abstract |
TEX101, a unique germ-cell-specific marker protein, shows sexually dimorphic expression during mouse gonad development. To clarify the molecular basis of TEX101, we performed the RNA interference (RNAi)-knockdown study of TEX101 expressed in mouse testis.
1. We examined the expression level of Tex101, by real-time PCR, in several cell lines derived from small cell carcinomas (e.g., Lu-139, and -140) and germ cell tumors (e.g., NEC-8 and -14) to find cell lines useful for in vitro analysis of TEX101. Among them, we found that Lu-140 expressed Tex101, albeit at a very weak level.
2. We generated short-hairpin RNA (shRNA) expression constructs against three target sites in Tex101 and used plasmids encoding the shRNA along with Green fluorescence protein (GFP), i.e., GFP-shTex101. We validated the inhibitory efficiency of the GFP-shTex101 vectors using Tex101-transfected COS-7 cells. The vectors highly inhibited the expression of Tex101 in the culture cells.
3. We next tried to transfect the GFP-shTex101 vectors into mouse testis using electroporation. Although GFP signal indicating transfection was detectable in germ cells in the testis, the transfection efficiency of the GFP-shTex101 vectors was poor. Detailed morphological changes on GFP-sh Tex101-transfeted mouse testis as well as improvement of in vivo transfection efficiency remain to be resolved.
4. We also investigated the biochemical and immunohistochemical characterization of TEX101 from mice.
GFP-shTex101 generated in this study would helpful in elucidating the molecular characteristics and its physiological functions of TEX101.
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Report
(3 results)
Research Products
(22 results)
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[Journal Article] MicroRNA cloning analysis reveals sex differences in microRNA expression profiles between adult mouse testis and ovary2008
Author(s)
Takuya Mishima, Takami Takizawa, Shan-shun Luo, Osamu Ishibashi, Yutaka Kawahigashi, Yoshiyuki Mizuguchi, Tomoko Ishikawa, Miki Mori, Tomohiro Kanda, Tadashi Goto, Takizawa Takizawa
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Journal Title
Reproduction 136(6)
Pages: 811-822
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Endothelial Expression of Fe Gamma Receptor IIb in the Full-term Human Placenta2007
Author(s)
Takuya Mishima, Gotaro Kurasawa, Gen Ishikawa, Miki Mori, Yutaka Kawahigashi, Tomoko Ishikawa, Shan-shun Luo, Takami Takizawa, Tadashi Goto, Shigeki Matsubara, Toshiyuki Takeshita, John M.Robinson, Toshihiro Takizawa
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Journal Title
Placenta 28(2-3)
Pages: 170-174
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Molecular characterization of a germ-cell-specific antigen, TEX101, from mouse testis2006
Author(s)
Hong Jin, Hiroshi Yoshitake, Hiroki Tsukamoto, Mai Takahashi, Miki Mori, Toshihiro Takizawa, Kenji Takamori, Hideoki Ogawa, Katsuyuki Kinoshita, Yoshihiko Araki
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Journal Title
Zygote 14(3)
Pages: 201-208
Description
「研究成果報告書概要(欧文)」より
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[Presentation] RNAi-knockdown analysis of a germ-cell-specific antigen, TEX101, in mouse testis2008
Author(s)
Toshihiro Takizawa, Takuji Kosuge, Akima Harada, Miki Mori, Osamu Ishibashi, Takami Takizawa, Yoshihiko Araki, Yoko Sato, Yoshitaka Hishikawa, Takehiko Koji
Organizer
49th Annual Meeting of Japanese Society for Histochemistry and Cytochemistry
Place of Presentation
Nagasaki, Japan
Year and Date
2008-10-05
Description
「研究成果報告書概要(欧文)」より
Related Report
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