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Introduction and evaluation of micro beads-assisted capillary electrophoresis for nucleic acid aptamer selection systems

Research Project

Project/Area Number 18H02002
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 34020:Analytical chemistry-related
Research InstitutionThe University of Tokyo

Principal Investigator

Yoshimoto Keitaro  東京大学, 大学院総合文化研究科, 准教授 (60392172)

Co-Investigator(Kenkyū-buntansha) 吉冨 徹  国立研究開発法人物質・材料研究機構, 機能性材料研究拠点, 主任研究員 (20585799)
齋藤 伸吾  埼玉大学, 理工学研究科, 教授 (60343018)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
Fiscal Year 2020: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2019: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Keywords核酸化学 / 分離化学 / 核酸アプタマー / キャピラリー電気泳動 / DNA / RNA / 蛍光 / バイオセンサー / 核酸 / 低分子化合物 / アプタマー / 分子進化工学 / 核酸科学 / 分子認識化学
Outline of Final Research Achievements

In this research project, the micro-particle-assisted capillary electrophoresis (MACE) system discovered by the applicant was introduced into a nucleic acid aptamer selection experiment (SELEX) system for various target molecules to demonstrate that there is still a large room for growth in the SELEX system, which has been attempted to be improved using a synthetic organic chemistry approach. The objective of this study was to demonstrate that the SELEX method can be significantly improved by using an "analytical chemistry" approach. As a result, we demonstrated that the MACE-SELEX method works on a wide range of targets, including soluble proteins, membrane proteins, and small molecules, and that it is possible to obtain DNA aptamers with high binding affinity.

Academic Significance and Societal Importance of the Research Achievements

核酸やペプチドを骨格とした分子認識能を有する生体高分子は、創薬分野での利用が期待されている中分子に分類される薬剤モダリティである。薬剤として高い潜在能力を有する候補中分子の獲得は、分子ライブラリーの中から、進化分子工学的手法により選抜する。従来有機合成化学的アプローチで改善が試みられていた SELEX 系に未だ大きな伸び代が存在すること、さらに“分析化学”的アプローチで SELEX 法の大幅な質の向上が可能であることを実証することを目的とした。

Report

(4 results)
  • 2021 Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (32 results)

All 2021 2020 2019 2018 Other

All Journal Article (8 results) (of which Peer Reviewed: 8 results,  Open Access: 5 results) Presentation (19 results) (of which Int'l Joint Research: 5 results,  Invited: 19 results) Remarks (3 results) Patent(Industrial Property Rights) (2 results) (of which Overseas: 1 results)

  • [Journal Article] Accelerated Discovery of Potent Bioactive anti‐TNFα Aptamers by Microbead‐Assisted Capillary Electrophoresis (MACE)‐SELEX2021

    • Author(s)
      Nagano Masanobu、Oguro Takumi、Sawada Ryo、Yoshitomi Toru、Yoshimoto Keitaro
    • Journal Title

      ChemBioChem

      Volume: 22 Issue: 23 Pages: 3341-3347

    • DOI

      10.1002/cbic.202100478

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Design Strategy of Short Antidote Sequence for Bivalent aptamer: Rapid Neutralization of High Anticoagulant Thrombin-Binding DNA Aptamer Linked M08 with HD222021

    • Author(s)
      Yoshitomi Toru、Wakui Koji、Miyakawa Masato、Yoshimoto Keitaro
    • Journal Title

      Research and Practice in Thrombosis and Haemostasis

      Volume: -

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Binding and Structural Properties of DNA Aptamers with VEGF-A-Mimic Activity2020

    • Author(s)
      Yoshitomi Toru、Hayashi Misako、Oguro Takumi、Kimura Keiko、Wayama Fumiya、Furusho Hitoshi、Yoshimoto Keitaro
    • Journal Title

      Molecular Therapy - Nucleic Acids

      Volume: 19 Pages: 1145-1152

    • DOI

      10.1016/j.omtn.2019.12.034

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] High Enrichment of Nucleobase-modified Aptamers in Early Selection Rounds by Microbeads-assisted Capillary Electrophoresis SELEX2019

    • Author(s)
      WAKUI Koji、ABE Akihito、YOSHITOMI Toru、FURUSHO Hitoshi、YOSHIMOTO Keitaro
    • Journal Title

      Analytical Sciences

      Volume: 35 Issue: 5 Pages: 585-588

    • DOI

      10.2116/analsci.18SDN04

    • NAID

      130007644236

    • ISSN
      0910-6340, 1348-2246
    • Year and Date
      2019-05-10
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Screening of DNA Signaling Aptamer from Multiple Candidates Obtained from SELEX with Next-generation Sequencing2019

    • Author(s)
      YOSHITOMI Toru、WAYAMA Fumiya、KIMURA Keiko、WAKUI Koji、FURUSHO Hitoshi、YOSHIMOTO Keitaro
    • Journal Title

      Analytical Sciences

      Volume: 35 Issue: 1 Pages: 113-116

    • DOI

      10.2116/analsci.18SDN05

    • NAID

      130007554959

    • ISSN
      0910-6340, 1348-2246
    • Year and Date
      2019-01-10
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Rapidly Neutralizable and Highly Anticoagulant Thrombin-Binding DNA Aptamer Discovered by MACE SELEX2019

    • Author(s)
      Wakui Koji、Yoshitomi Toru、Yamaguchi Akane、Tsuchida Maho、Saito Shingo、Shibukawa Masami、Furusho Hitoshi、Yoshimoto Keitaro
    • Journal Title

      Molecular Therapy - Nucleic Acids

      Volume: 16 Pages: 348-359

    • DOI

      10.1016/j.omtn.2019.03.002

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Programmable RNA detection with a fluorescent RNA aptamer using optimized three-way junction formation2019

    • Author(s)
      Furuhata Yuichi、Kobayashi Mizuki、Maruyama Ryo、Sato Yusuke、Makino Kurumi、Michiue Tatsuo、Yui Hiroharu、Nishizawa Seiichi、Yoshimoto Keitaro
    • Journal Title

      RNA

      Volume: 25 Issue: 5 Pages: 590-599

    • DOI

      10.1261/rna.069062.118

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Estimation of G-quartet-forming guanines in parallel-type G-quadruplexes by optical spectroscopy measurements of their single-nucleobase substitution sequences2018

    • Author(s)
      Maruyama Ryo、Makino Kurumi、Yoshitomi Toru、Yui Hiroharu、Furusho Hitoshi、Yoshimoto Keitaro
    • Journal Title

      The Analyst

      Volume: 143 Issue: 17 Pages: 4022-4026

    • DOI

      10.1039/c8an01131a

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Presentation] 新規な分離技術を導入した分子進化工学的手法 MACE-SELEX で獲得した DNA アプタマーの機能2021

    • Author(s)
      吉本敬太郎
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Reguration of Cellar Functions by Nucleic Acid Aptamers2021

    • Author(s)
      吉本敬太郎
    • Organizer
      Pacifichem 2021
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 新規な分離科学を取り入れた分子進化工学的手法の構築 -同手法で獲得したアンタゴニスト型核酸分子の機能-2020

    • Author(s)
      吉本敬太郎
    • Organizer
      第43回日本分子生物学会年会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 血液凝固因子を標的とする中和可能な核酸アプタマーの設計2020

    • Author(s)
      吉本敬太郎
    • Organizer
      第69回高分子討論会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] Anti-coagulant activity of DNA aptamer and its neutralization by antidote single stranded DNA2020

    • Author(s)
      吉本敬太郎
    • Organizer
      第42回日本血栓止血学会学術集会
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Rapidly Neutralizable and High Anticoagulant DNA Aptamer Discovered by MACE-SELEX2020

    • Author(s)
      吉本敬太郎
    • Organizer
      Pittcon 2020
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 分子認識能をもつ核酸“核酸アプタマー”の医療・創薬分野への挑戦2020

    • Author(s)
      吉本敬太郎
    • Organizer
      九州大学先導物質化学研究所講演会
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 新規な分離科学を取り入れた分子進化工学的手法の構築 -同手法で獲得したアンタゴニスト型核酸分子の機能-2020

    • Author(s)
      吉本 敬太郎
    • Organizer
      第43回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 血液凝固因子を標的とする中和可能な核酸アプタマーの設計2020

    • Author(s)
      吉本 敬太郎
    • Organizer
      第69回高分子討論会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] Anti-coagulant activity of DNA aptamer and its neutralization by antidote single stranded DNA2020

    • Author(s)
      吉本 敬太郎
    • Organizer
      第42回日本血栓止血学会学術集会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] toehold 導入型核酸アプタマーによる中和制御可能な抗凝固製剤2019

    • Author(s)
      吉本敬太郎
    • Organizer
      日本薬学会第139年会 一般シンポジウム “生体分子骨格リデザインで挑む次世代創薬研究”, 2019年3月22日, (幕張メッセ, 幕張, 千葉)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] MACE/NGS-SELEXで発見されたトロンビンアプタマーの抗凝固活性と中和2019

    • Author(s)
      吉本敬太郎
    • Organizer
      日本化学会第99春季年会特別企画 “核酸分析・機能創出の新展開”, 2019年3月19日, (甲南大学, 神戸, 兵庫)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] Functions of DNA aptamers discovered by improved SELEX2019

    • Author(s)
      吉本敬太郎
    • Organizer
      3rd International symposium on Nanoarchitectonics for Mechanobiology, March 8th 2019 (NIMS, Tsukuba, Ibaraki)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] マイクロ粒子支援型キャピラリー電気泳動 SELEX で獲得した抗トロンビンアプタマーの抗凝固能2018

    • Author(s)
      吉本敬太郎
    • Organizer
      第38回 キャピラリー電気泳動シンポジウム (SCE2018), 2018年12月6日(I-siteなんば 大阪府立大学, 大阪, 大阪)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 分子標的薬のスクリーニング実験における分離・洗浄工程の重要性 ~ 核酸アプタマースクリーニングを例に~2018

    • Author(s)
      吉本敬太郎
    • Organizer
      甲南大学 Science Live Ticket Series Part 36, 2018年11月20日(甲南大学, 神戸, 兵庫)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] DNAアプタマー選抜における分離・洗浄工程の重要性:微粒子導入型キャピラリー電気泳動の導入効果2018

    • Author(s)
      吉本敬太郎
    • Organizer
      日本分析化学会第67年会, 2018年7月30日(東京大学生産技術研究所, 駒場, 東京)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 分子認識型核酸(核酸アプタマー)選抜実験における分離・洗浄工程の重要性2018

    • Author(s)
      吉本敬太郎
    • Organizer
      平成30年度第1回奨励会特別研究会, 2018年7月30日(東京大学生産技術研究所, 駒場, 東京)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 核酸アプタマー選抜実験系(SELEX)における分離・洗浄工程の重要性2018

    • Author(s)
      吉本敬太郎
    • Organizer
      第2回中分子×IT×創薬ビジネス研究会, 2018年5月31日(川崎市産業振興会館, 川崎, 神奈川)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] MACE-SELEX により選抜された抗トロンビン DNA アプタマーの抗凝固作用2018

    • Author(s)
      吉本敬太郎
    • Organizer
      バイオメディカル研究部門セミナー, 2018年1月30日(産業技術総合研究所, つくば, 茨城)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Remarks] リンクバイオ、薬剤候補スクリーニングを「視える化」し効率アップ

    • URL

      https://bio.nikkeibp.co.jp/atcl/column/16/021500017/042100238/

    • Related Report
      2020 Annual Research Report
  • [Remarks] Publication List (吉本研究室)

    • URL

      http://yoshimotolab.c.u-tokyo.ac.jp/publication/

    • Related Report
      2018 Annual Research Report
  • [Remarks] Patent & Press (吉本研究室)

    • URL

      http://yoshimotolab.c.u-tokyo.ac.jp/patent-and-press/

    • Related Report
      2018 Annual Research Report
  • [Patent(Industrial Property Rights)] 核酸アプタマー2019

    • Inventor(s)
      吉本、吉冨、和久井、古性、齋藤、宮内
    • Industrial Property Rights Holder
      吉本、吉冨、和久井、古性、齋藤、宮内
    • Industrial Property Rights Type
      特許
    • Filing Date
      2019
    • Related Report
      2018 Annual Research Report
    • Overseas
  • [Patent(Industrial Property Rights)] 解毒剤2018

    • Inventor(s)
      吉本、吉冨、和久井、古性
    • Industrial Property Rights Holder
      吉本、吉冨、和久井、古性
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2018-231878
    • Filing Date
      2018
    • Related Report
      2018 Annual Research Report

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Published: 2018-04-23   Modified: 2023-01-30  

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