Establishment of Neu1 knockout zebrafish by genome-editing
| Project/Area Number |
16K07876
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Aquatic life science
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| Research Institution | Kagoshima University |
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Principal Investigator |
Shiozaki Kazuhiro 鹿児島大学, 農水産獣医学域水産学系, 准教授 (70390896)
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2016: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
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| Keywords | シアリダーゼ / リソソーム / ゲノム編集 / シアル酸 / ゼブラフィッシュ / 糖鎖 / メダカ / モデル動物 |
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| Outline of Final Research Achievements |
Mammalian sialidases have been studied so far. Among them, lysosomal sialidases are known to be involved in immunity, autophasy and sialidosis. On the other hand, the functions of fish lysosomal sialidase is not clear. Here, we tried to clarify the significance of fish lysosomal sialidase using gene-modified fish.
First, Neu1 polypeptide knockdown using morpholino-oligo was conducted in medaka embryo. Knockdown embryo exhibited the accumulation of sialoglycoprotein and abnormal phenotypes such as higher heart rate. Enzymatic properties of fish Neu1 are found to be highly conserved by gene cloning and polypeptide analysis. Next, we tried to establish Neu1 knockout zebrafish by CRISPR/Cas9 method. Neu1 knockout zebrafish showed alteration of muscle- and bone-related gene expressions in embryo accompanied with the decrease of sialidase activity. Adult fish showed the abnormal bone shape. As a result, fish Neu1 would be crucial in bone formations.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では魚類Neu1シアリダーゼの機能について明らかにしたが、この表現型はヒトNeu1欠損症であるシアリドーシスの症状と非常に類似したものがあった。シアリドーシスは胎児期に進行する疾病であり、また希少疾患であることからその発症メカニズムや病気の進行に関する知見は非常に限られている。本研究で用いたゼブラフィッシュは多産であり、しかも卵や孵化後の仔魚は透明であることからヒト胎児の研究に非常に優れた実験動物といえる。今後Neu1ノックアウトゼブラフィッシュがシアリドーシス疾病モデルとして利用できる可能性がある。
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Report
(4 results)
Research Products
(23 results)
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[Journal Article] Adjuvant effect of recombinant interleukin-12 in the Nocardiosis formalin-killed vaccine of the amberjack Seriola dumerili2017
Author(s)
Matsumoto, M., Araki, K., Hayashi, K., Takeuchi, Y., Shiozaki, K., Suetake, H., Yamamoto, A
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Journal Title
Fish Shellfish Immunol
Volume: 67
Pages: 263-269
DOI
Related Report
Peer Reviewed
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[Presentation] Characterization of tilapia sialidases and its significance in bacterial infection2017
Author(s)
Honda, A., Chigwechokha, KP., Wakamatsu, R., Kamada, Y., Komatsu, M., Shiozaki, K.
Organizer
The JSFS 85th Anniversary-Commemorative International Symposium“Fisheries Science for Future Generations”,
Related Report
Int'l Joint Research
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