Development of antibody-fusion proximity biotinylation enzyme for analysis of virus entry
| Project/Area Number |
21K19230
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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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| Review Section |
Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
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| Research Institution | Ehime University |
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Principal Investigator |
Sawasaki Tatsuya 愛媛大学, プロテオサイエンスセンター, 教授 (50314969)
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| Project Period (FY) |
2021-07-09 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2021: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
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| Keywords | 近位依存性ビオチン標識法 / ウイルス / 感染症 / AirID / ビオチン / 新型コロナウイルス / レンチウイルス / 近位依存性ビオチン化酵素 / 抗体 / SARS-CoV-2 |
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| Outline of Research at the Start |
本申請課題はレンチウイルスをモデルに、ウイルス認識抗体FabにAirIDが融合した抗ウイルス-AirIDの構築し、抗体-AirIDを付与されたウイルスの作製を行う。構築した抗体-AirIDがウイルス表面タンパク質をビオチン標識できるかどうか検証を行う。さらに、抗体-AirIDが付加されたウイルスの細胞への感染能を確認した後、抗体-AirID付与ウイルス感染細胞において、質量分析によりビオチン化されるヒト宿主タンパク質の同定を行う。
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| Outline of Final Research Achievements |
Using lentivirus as a model, we applied the proximal-dependent biotinyl labeling method to elucidate the virus infection entry pathway, consisting of 1) construction of an anti-virus-AirID fused with a virus recognition antibody Fab, 2) generation of antibody-AirID-loaded virus, 3) verification of biotinylation of virus surface proteins, 4) confirmation of infection ability in cells, and 5) identification of antibody-AirID-loaded We developed technologies for the following five research projects: 1) creation of biotinylated viruses, 2) creation of biotinylated viruses, 3) validation of biotinylation of viral surface proteins, 4) confirmation of virus infectivity, and 5) identification of biotinylated host proteins in cells infected with virus. With this method, the proximal-dependent biotinyl labeling method was applied to elucidate the viral infection entry pathway.
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| Academic Significance and Societal Importance of the Research Achievements |
ウイルスの構造タンパク質を特異的に認識する抗体に近位依存性ビオチン化酵素を融合することにより、ウイルス粒子を近位依存性ビオチン化酵素でラベルし、ウイルスが細胞へ侵入するために関与する宿主タンパク質をビオチン化という形で検出・同定する技術の開発は、世界初の試みであるため挑戦的で社会的意義は大きい。
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Report
(3 results)
Research Products
(40 results)
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[Journal Article] Immune gene activation by NPR and TGA transcriptional regulators in the model monocot Brachypodium distachyon2022
Author(s)
Shimizu K., Suzuki H., Uemura T., Nozawa A., Desaki Y., Hoshino R., Yoshida A., Abe H., Nishiyama M., Nishiyama C., Sawasaki T., Arimura G.
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Journal Title
Plant Journal
Volume: 10
Issue: 2
Pages: 470-481
DOI
Related Report
Peer Reviewed
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[Journal Article] Myofiber androgen receptor increases muscle strength mediated by a skeletal muscle splicing variant of Mylk42021
Author(s)
Iori Sakakibara, Yuta Yanagihara, Koichi Himori, Takashi Yamada, Hiroshi Sakai, Yuichiro Sawada, Hirotaka Takahashi, Noritaka Saeki, Hiroyuki Hirakawa, Atsushi Yokoyama, So-Ichiro Fukada, Tatsuya Sawasaki, Yuuki Imai
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Journal Title
iScience
Volume: 24
Issue: 4
Pages: 102303-102303
DOI
Related Report
Peer Reviewed / Open Access
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