| Project/Area Number |
25660257
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Integrative animal science
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| Research Institution | Tokyo Denki University |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | バイオテクノロジー / 前核間細胞質マイクロインジェクション / 遺伝子 / 人工染色体ベクター / 高濃度 / 生存率 / 発現率 / 振動型マイクロインジェクション法 / マイクロインジェクショ / マイクロインジェクション |
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| Outline of Final Research Achievements |
Out of two new gene-transferring methods, Inter-pronuclear Cytoplasmic Microinjection (IPCM) showed good results. IPCM delivers a solution containing a high concentration of large transgene into a narrow space between two pronuclei and thereby never causes such a fatal event that a micropipette pulls out pronuclear components. We made solutions of 300-kb BAC DNA with tdTomato at 4 concentrations of 2, 5, 7.5 and 10 ng/uL, and tested their respective effects in 1,914 murine zygotes. The rates of embryos developing to the blastocyst stage were 79.7 (392/492), 64.3 (261/406), 72.9 (291/399) and 54.0% (333/617) as the concentration increased, but the 7.5- and 10-ng/uL groups not only showed significantly better rates of blastocysts expressing tdTomato than the others, but also attained the rates as high as 35.8 (143/399) and 39.9% (246/617), respectively. These results suggest that IPCM is a promising gene transfer that can be expected to achieve a high efficiency of genetic modification.
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