| Project/Area Number |
13358014
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Laboratory animal science
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| Research Institution | Shiga university of medical science, Research center for animal life science |
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Principal Investigator |
TAKADA Tatsuyuki Shiga university of medical science, Research center for animal life science, Associate professor, 動物生命科学研究センター, 助教授 (90206756)
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| Co-Investigator(Kenkyū-buntansha) |
TORII Ryuzo Shiga university of medical science, Research center for animal life science, Professor, 動物生命科学研究センター, 教授 (50106647)
TSUCHIYA Hideaki Shiga university of medical science, Research center for animal life science, Research assistant, 動物生命科学研究センター, 助手 (10378440)
KIMURA Hiroshi Shiga university of medical science, Department of experimental radiology, Professor, 医学部, 教授 (00110560)
KIMOTO Yasuhiko Tanabe seiyaku Co., Ltd., Discovery research laboratory, Director, 創薬研究所, 所長
細井 美彦 近畿大学, 生物理工学部, 助教授 (70192739)
入谷 明 近畿大学, 生物理工学部, 教授 (80026385)
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| Project Period (FY) |
2001 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥49,920,000 (Direct Cost: ¥38,400,000、Indirect Cost: ¥11,520,000)
Fiscal Year 2004: ¥10,140,000 (Direct Cost: ¥7,800,000、Indirect Cost: ¥2,340,000)
Fiscal Year 2003: ¥9,880,000 (Direct Cost: ¥7,600,000、Indirect Cost: ¥2,280,000)
Fiscal Year 2002: ¥11,960,000 (Direct Cost: ¥9,200,000、Indirect Cost: ¥2,760,000)
Fiscal Year 2001: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000)
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| Keywords | monkey ES cell / GFP / genetic engineering / nuclear transfer / ES細胞 / 遺伝子導入 / 多能性 / 分化 |
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| Research Abstract |
There are two major objectives in this research project. One is the genetic engineering of monkey ES cells and the other is genetic engineering of monkey using ES cells. In the first objective, we have succeeded to establish the monkey Es cell line which express GFP and showed that this cell line had pluripotency in vitro and in vivo. In addition, we established the efficient gene silencing method in monkey ES cells using siRNA. Therefore we were able to establish the method that allows over-expression and repression in monkey ES cells.
With regard to the second objective, we injected these ES cells into four-cell stage preimplantation embryos and found that these ES cells proliferated in the embryo to form chimeric blastocysts, suggesting this ES cell line has developmental potency. Then we proceeded to transfer these chimeric embryos to the oviduct of the recipient monkey. We succeeded to give pregnancy in three cases and had healthy birth of one infant. We analyzed the GFP expression in skin and blood cells under fluorescent light, but no obvious GFP expression was observed. However, when we analyzed the genomic DNA for the presence of GFP gene by PCR method, we were able to detect the GFP gene. These results strongly suggest that we succeeded to produce chimeric monkey for the first time. Now we are investigating GFP expression and chimeric ratio in detail.
Application of this technology enables the production of genetically engineered monkeys which are useful for disease models.
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