| Project/Area Number |
17K19398
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biology of Cells to Organisms, and related fields
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| Research Institution | Keio University |
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Principal Investigator |
Akanuma Takashi 慶應義塾大学, 医学部(信濃町), 助教 (50450721)
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| Project Period (FY) |
2017-06-30 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2017: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
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| Keywords | ゼブラフィッシュ / ライブイメージング / 運命決定 / ゲノム編集 / ノックイン / 発生・分化 / 定量解析 / 形 / 非遺伝的 |
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| Outline of Final Research Achievements |
In this study, I first developed a simple and rapid method of homologous recombination-based gene targeting to generate genetically modified zebrafish in which fluorescent proteins are expressed in a cell type-specific manner. Injection of the donor DNA comprising fluorescent protein fragments and long (~3kb) homology arms together with chromatin modifier inhibitors into fertilized zebrafish eggs results in efficient targeting at the predetermined genomic location. Using my novel gene targeting method, I created transgenic zebrafish which expresses the membrane-bound GFP protein in V2 interneuron progenitors (V2 cells). Through the imaging analysis of this zebrafish, I succeeded in recording the V2 cell behaviors over a long time and identified characteristic features of V2 cell shape change. Now I am investigating the relationship between the cell shape change history and fate determination of daughter cells after the asymmetric V2 cell division.
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| Academic Significance and Societal Importance of the Research Achievements |
CRISPR-Cas9システムを用いたゲノム編集は、近年盛んに行われているが、ゼブラフィッシュでは効率の面で課題があった。これに対し本研究課題の成果であるCRISPR-Cas9システムを使わない遺伝子改変技術により、従来よりも簡便に遺伝子改変ゼブラフィッシュを作製できるようになると思われる。そして、ゼブラフィッシュモデルの特長であるイメージング解析にとって有用なトランスジェニックラインが今まで以上に作られるようになり、発生学のみならず様々な分野で活用されることが期待される。
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