Water-in-oil method for single-cell proteomics
Project/Area Number |
19K05544
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Review Section |
Basic Section 34020:Analytical chemistry-related
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Research Institution | Kumamoto University |
Principal Investigator |
Masuda Takeshi 熊本大学, 大学院生命科学研究部(薬), 助教 (70383940)
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Co-Investigator(Kenkyū-buntansha) |
宇都 甲一郎 国立研究開発法人物質・材料研究機構, 国際ナノアーキテクトニクス研究拠点, 独立研究者 (30597034)
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Project Period (FY) |
2019-04-01 – 2022-03-31
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Project Status |
Completed (Fiscal Year 2021)
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Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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Keywords | プロテオミクス / 高感度分析 / 1細胞オミクス / 油中液滴 |
Outline of Research at the Start |
1細胞から1,000種類以上のタンパク質をハイスループットに定性・定量できる分析システムを開発する。1細胞プロテオミクスを達成することで、同一細胞群内における分子レベルの不均一性発生機構や不均一性が生み出す発生や分化に関わる複雑な生命機構の解明につながる。これまでにない分解能および視点から生命活動を観察できる。目的を達成するために、申請者が独自開発した油中液滴チャンバー法を基盤とする。
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Outline of Final Research Achievements |
In this project, we aim to establish a practical single-cell proteomics system. This study was based on the water droplet-in-oil digestion (WinO) method which was established by the applicant. We developed a pseudo amplification method for proteins using stable isotope-labeled reagents. To achieve high throughput analysis, we established a semi-automated pretreatment using a liquid handling robot. These improvements dramatically increased the number of quantified proteins in single-cell proteomics. We also developed a superhydrophobic materials suitable for the WinO method.
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Academic Significance and Societal Importance of the Research Achievements |
細胞の分子レベルの不均一性は、組織や器官の複雑な機能やガン細胞集団としての抗ガン剤耐性を生み出すなど、重要な生命現象に深く関与している。本申請課題では、これまで難しかった1細胞プロテオミクスの実現に大きく近づける基盤技術を構築した。1細胞プロテオミクスを実現することで、従来のプロテオミクスでは見落としていた新たな現象の発見につながることで、学術的意義が大きい成果と言える。また、1細胞プロテオミクス技術は新規治療標的の探索などへの応用にも期待できることから、本成果は社会的意義もある。
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Report
(4 results)
Research Products
(49 results)
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[Journal Article] Lysine Demethylase 5A is Required for MYC Driven Transcription in Multiple Myeloma2021
Author(s)
Ohguchi H、Park Paul M.C.、wang t、Gryder B E.、Ogiya D、kurata k、zhang x、Li D、pei c、Masuda T、Johansson C、Wimalasena V K、Kim Y、Hino S、Usuki S、Kawano Y、Samur M K、Tai Yu-Tzu、Munshi N C.、Matsuoka M、Ohtsuki S、Nakao M、Minami T、Lauberth S、Khan J、Oppermann U、Durbin A D.、Anderson K C.、Hideshima T、Qi J
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Journal Title
Blood Cancer Discovery
Volume: in press
Issue: 4
Pages: 370-387
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] Cyclocreatine Transport by SLC6A8, the Creatine Transporter, in HEK293 Cells, a Human Blood-Brain Barrier Model Cell, and CCDSs Patient-Derived Fibroblasts.2020
Author(s)
Uemura T, Ito S, Masuda T, Shimbo H, Goto T, Osaka H, Wada T, Couraud PO, Ohtsuki S.
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Journal Title
Pharm Res.
Volume: 37
Issue: 3
Pages: 1-11
DOI
Related Report
Peer Reviewed
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[Journal Article] Large-Scale Quantitative Comparison of Plasma Transmembrane Proteins between Two Human Blood-Brain Barrier Model Cell Lines, hCMEC/D3 and HBMEC/cibeta.2019
Author(s)
Masuda T, Hoshiyama T, Uemura T, Hirayama-Kurogi M, Ogata S, Furukawa A, Couraud PO, Furihata T, Ito S, Ohtsuki S
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Journal Title
Mol Pharm
Volume: 16
Issue: 5
Pages: 2162-2171
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Presentation] Cyclocreatine transport by creatine transporter (SLC6A8) in HEK293 cells, a human blood-brain barrier model cell, and CCDSs patient-derived fibroblasts2021
Author(s)
Shingo Ito, Tatsuki Uemura, Takeshi Masuda, Hiroko Shimbo, Tomohide Goto, Hitoshi Osaka, Takahito Wada, Pierre-Olivier Couraud, Sumio Ohtsuki
Organizer
36th JSSX Annual Meeting
Related Report
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