| Project/Area Number |
14580798
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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 |
Laboratory animal science
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| Research Institution | Shiga university of medical 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) |
KIMURA Hiroshi Shiga university of medical science, Department of experimental radiology, Professor, 医学部, 教授 (00110560)
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)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | RNA interference / knock out / siRNA / ES細胞 / ノックダウン / ARL |
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| Research Abstract |
We initially tried to introduce siRNA into fertilized embryos and blastocysts. But it was not efficient. Then we decided to use ES cells as they were developmentally similar stage to blastocysts and able to contribute to entire animal cells. First, we established GFP expressing mouse and monkey EScells and tried to suppress GFP, because suppression of GFP can be easily monitored by fluorescence. We succeeded to suppress GFP expression using GFP specific siRNA efficiently. Suppression was confirmed on both mRNA and protein level. Furthermore, Oct4 which plays important role for the maintenance of pluripotency was effectively suppressed with this method, and as a result, ES cells differentiated into trophectoderm cells, suggesting this method can be used as a novel differentiation method of ES cells.
We demonstrated that siRNA is useful to suppress specific gene for a specific period in ES cells. Using this method, stage-specific knock out in mouse can be achieved by making chimeric blastocysts consisting of ES cells which were introduced siRNA and tetraploid blastocysts. This method is useful not only for stage-specific suppression of gene but also differentiation of ES cells.
The character that siRNA is not integrated into genome and do not modify genome organization is ideal for stem cell therapy.
Our result should contribute to the developmental biotechnology and regenerative medicine.
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