| Project/Area Number |
17300135
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Osaka University |
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Principal Investigator |
OKABE Masaru Osaka University, Research Institute for Microbial Diseases, Professor (30089875)
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| Co-Investigator(Kenkyū-buntansha) |
IKAWA Masahito Osaka University, Research Institute for Micaobial Diseases, Associate Professor (20304066)
INOUE Naokazu Osaka University, Research Institute for Microbial Diseases, Assistant Professor (50379096)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥15,950,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2007: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
Fiscal Year 2006: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 2005: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Keywords | bioima ging / placenta specific gene manipulation / lentivirus / implantation / giant cells / fusion / syncytin / レンチウィルス / GFP / 精子 / 卵子 / 受精 / 胎盤 / 遺伝子導入 / 蛍光たんぱく質 / 子宮 / 輸卵管 |
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| Research Abstract |
The placenta plays important roles to support fetal development, such as gas exchange, nutrient supply, and hormone production. Placental defects underlie many aspects of pregnancy losses and complications ; thus, understanding and regulating gene function during placentation is of high clinical relevance. However, the lack of a facile and efficient method for placenta-specific gene manipulation has hampered the study of placenta. During to find the way to visualize the placentation, we tried to apply the lentivirus (LV) vectors to organ specific gene manipulation. We have previously shown that the transduction of fertilized mouse eggs with LV vectors efficiently generates transgenic animals, however transgene expression occurred in both the fetus and the placenta. In our study performed under the support of this grant, we transduced zona-free blastocysts with LV vectors expecting placenta-specific gene expression, since most placental cells differentiate from trophoblast cells that fo
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rm the outermost layer of the blastocyst. Since we used GFP as a transgene, the transgene expression was observed by fluorescent microscopes. As a result, the GFP fluorescence was found in trophoblast cells from preimplantation stages and in placenta throughout gestation. All the analyzed placentas carried the transgene, while none of the fetuses became transgenic. Thus to establish the basic method to visualize the placenta was achieved. In order to develop the application use, we further performed experiments. For the application of this method, embryonic lethal mouse strains were chosen in which the embryos die by placental defects. In our experiment, we could demonstrate that several knockout animal models were substantially rescued by the introduction of transgene exclusively into the placenta. This technology provides a powerful system for gene manipulation exclusively in placental organogenesis with implications for future clinical usage for the gene therapy for placental dysfunction. Less
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