| 研究課題/領域番号 |
16F16416
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| 研究機関 | 東京大学 |
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研究代表者 |
河岡 義裕 東京大学, 医科学研究所, 教授 (70135838)
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| 研究分担者 |
WANG I-HSUAN 東京大学, 医科学研究所, 外国人特別研究員
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| 研究期間 (年度) |
2016-11-07 – 2019-03-31
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| キーワード | Influenza virus / Viral genome packaging / Virus-host interaction / Single-molecule FISH |
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| 研究実績の概要 |
Influenza viruses are a common cause of human respiratory infections, which lead to seasonal, endemic infectious disease every year. Influenza A virus (IAV) is an enveloped virus with eight segments of distinct single-stranded, negative-sense RNA as genomic material. One of the essential processes to replicate IAV is to correctly package its fragmented genome into progeny virions. To understand the mechanism of this critical step, we proposed to systematically identify and study the cellular factors that contribute to the genome packaging of IAV. In the first year of this project, we developed image-based assays that allow us to directly visualize and monitor viral RNA segments in infected cells. Compared to the traditional molecular virology methods, the quantitative microscopy approach grants better temporal and spatial resolution to the assays. By analyzing the properties of viral RNA in the images, we can now inspect individual viral RNA segments at specific steps during infection. We also employed the established assays to screen for the cellular protein(s) involved in viral genome packaging. Our goals are to pinpoint the key factor(s) in the scenario, and investigate how viral genome packaging is carried out by the virus and host factor(s). We believe that this work will lead to a comprehensive understanding of the fundamental biology of influenza virus.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
1: 当初の計画以上に進展している
理由
We have planned to develop a single-cell based quantitative method to monitor viral RNA during the infection in the first year. Now, an assay that combines single-molecule fluorescence in situ hybridization and computational image analyses is set up to track individual viral RNA segments and determine the kinetics of multiple events at the late stage of infection. We further automated the established analyses for use in a planned siRNA screen. The screen, which was originally scheduled in the second year, already began and is halfway through.
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| 今後の研究の推進方策 |
As a next step, we will complete the ongoing siRNA screen, which aims to verify the 28 potential host targets identified in our previous interactomic study of influenza virus infection. We hope that we can proceed to the mechanistic study as soon as the promising host factor involved in viral genome packaging is found. In addition to the screen, we will also exploits the same method to study the temporal and spatial coordination of the inter-segment interaction between viral RNAs.
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