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Development of novel microRNA detection system

Research Project

Project/Area Number 16K14027
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Analytical chemistry
Research InstitutionNational Defense Medical College

Principal Investigator

TAKEI FUMIE  防衛医科大学校(医学教育部医学科進学課程及び専門課程、動物実験施設、共同利用研究施設、病院並びに防衛, 医学教育部, 准教授 (30252711)

Co-Investigator(Renkei-kenkyūsha) NAKATANI Kazuhiko  大阪大学, 産業科学研究所, 教授 (70237303)
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
KeywordsPCR / reverse transcription / fluorescence molecule / miRNA / 蛍光分子 / マイクロRNA / RT-PCR / ヘアピンプローブPCR / HP-PCR / バイオテクノロジー / 有機化学 / 遺伝子
Outline of Final Research Achievements

A novel rapid, simple, and cost-effective miRNA detection system using a primer having a tag at the 5' end was developed. miRNA is the 20-25 mers, and it is too short RNA to do reverse transcription (RT)-polymerase chain reaction (PCR) under the general RT-PCR method. We used a RT-primer having DNA tag to increase the length of cDNA. In PCR, tag-primer with template cDNA having complementary sequence of tag hybridized, and melting temperature of primer and cDNA increased. As a result, PCR efficiency increased, and miRNA can be detected quantatively.

Academic Significance and Societal Importance of the Research Achievements

microRNA(miRNA)は21-25塩基長の1本鎖RNA分子であり、真核生物において遺伝子の転写後発現調節に関与する。現在ヒトのmiRNAは2500種類以上知られており、様々な疾患や病気に関わっていることが明らかにされている。本提案では、PCRを基盤とした簡便、高感度なmiRNA検出法の開発を目的とし、研究を行なった結果、特殊な技術、試薬がなくても、誰もがmiRNAを検出できる画期的な方法の開発に成功した。今後、miRNAの検出分野において大きな役割を果たすと考えられる。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (15 results)

All 2018 2017 2016

All Journal Article (3 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 3 results) Presentation (11 results) (of which Int'l Joint Research: 7 results,  Invited: 1 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] PCR under Low Ionic Concentration Buffer Conditions2018

    • Author(s)
      Takei, F. ; Akiyama, M. ; Nobusawa, K. ; Sabani, Norhayati Binti ; Han, H. ; Nakatani, K. ; Yamashita, I.
    • Journal Title

      Chemistryselect

      Volume: 3 Issue: 3 Pages: 973-976

    • DOI

      10.1002/slct.201702542

    • Related Report
      2018 Annual Research Report 2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Fluorescence turn-on hairpin-probe PCR2018

    • Author(s)
      Takei, F.; Nakatani, K.
    • Journal Title

      Chem. Commun.

      Volume: 53 Issue: 8 Pages: 1393-1396

    • DOI

      10.1039/c6cc08947j

    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Synthesis and photo-physical properties of fluorescence molecular probe for turn-ON type detection of cytosine bulge DNA2016

    • Author(s)
      Verma, R.; Takei, F.; Nakatani, K.
    • Journal Title

      Org. Lett.

      Volume: 18 Issue: 13 Pages: 3170-3173

    • DOI

      10.1021/acs.orglett.6b01378

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] Development of novel miRNA detection system using PCR with C-Bulge probe and fluorescence molecule2018

    • Author(s)
      Fumie Takei, Misaki Akiyama, Asako Murata,Akiko Sugai, Kazuhiko Nakatani, Ichiro Yamashita
    • Organizer
      第45回国際核酸化学シンポジウム(ISNAC 2018)/日本核酸化学会第2回年会 ISNAC2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] A novel miRNA detection system using PCR with DNA probe and fluorescence molecule.2018

    • Author(s)
      Fumie Takei, Misaki Akiyama, Asako Murata,Akiko Sugai, Kazuhiko Nakatani, Ichiro Yamashita
    • Organizer
      IWSBN2018
    • Related Report
      2018 Annual Research Report
  • [Presentation] 低塩濃度におけるアレル特異的PCR法の開発2017

    • Author(s)
      武井史恵,秋山美咲,信澤和幸, Norhayati Binti Sabani, 韓 煥文, 中谷和彦, 山下一郎
    • Organizer
      第64回応用物理学会春季学術講演会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2017-03-14
    • Related Report
      2016 Research-status Report
  • [Presentation] Polymerase Chain Reaction (PCR) under low ionic strength2017

    • Author(s)
      H. Han, F. Takei, M. Akiyama, K. Nobusawa, N. B. Sabani, K. Nakatani, I. Yamashita
    • Organizer
      第64回応用物理学会春季学術講演会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2017-03-14
    • Related Report
      2016 Research-status Report
  • [Presentation] The PCR amplification limit under low ionic conditions2017

    • Author(s)
      Huan-wen Han,Kazuyuki Nobusawa, Fumie Takei, Norhayati Binti Sabani, Ichiro Yamashita, Kazuhiko Nakatani
    • Organizer
      9th International Conference on Molecular Electronics and Bioelectronics (M&BE9)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] PCR primer detection via molecular recognition by surface modified molecular ligand2017

    • Author(s)
      Kazuyuki Nobusawa, Fumie Takei, Norhayati Binti Sabani, Huan-wen Han, Ichiro Yamashita, Kazuhiko Nakatani
    • Organizer
      9th International Conference on Molecular Electronics and Bioelectronics (M&BE9)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Development of a SNP typing in the low ionic PCR condition2017

    • Author(s)
      Fumie Takei, Misaki Akiyama, Kazuyuki Nobusawa, Norhayati Binti Sabani, Huanwen Han, Kazuhiko Nakatani, Ichiro Yamashita
    • Organizer
      9th International Conference on Molecular Electronics and Bioelectronics (M&BE9)
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] A new idea of PCR progress monitoring by Electrochemical Impedance Spectroscopy (EIS): combination of ligands and hairpin primer2017

    • Author(s)
      Huanwen Han, Norhayati Binti Sabani, Kazuyuki Nobusawa, Fumie Takei, Kazuhiko Nakatani, Ichiro Yamashita
    • Organizer
      IWSBN2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] Design of ligand for sensing: the combination of hairpin primer DANP ligand.2017

    • Author(s)
      Fumie Takei, Misaki Akiyama, Kazuyuki Nobusawa, Norhayati Binti Sabani, Kazuhiko Nakatani, Ichiro Yamashita,
    • Organizer
      IWSBN2017
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] 1つの蛍光分子とDNAの特殊構造によって開発されたDNA増幅反応2017

    • Author(s)
      武井史恵
    • Organizer
      第30回産研技術室講演会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] Development of the allele specific PCR under the low ion PCR condition,2 Kazuhiko Nakatani,2 Ichiro Yamashita2016

    • Author(s)
      F. Takei, M. Akiyama, K. Nobusawa, N. B. Sabani, K. Nakatani, I. Yamashita
    • Organizer
      The 43rd International Symposium on Nucleic Acids Chemistry
    • Place of Presentation
      熊本大学
    • Year and Date
      2016-09-27
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Patent(Industrial Property Rights)] miRNA用プライマー及び該プライマーを用いた核酸増幅反応法2018

    • Inventor(s)
      武井 史恵
    • Industrial Property Rights Holder
      武井 史恵
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Related Report
      2018 Annual Research Report

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Published: 2016-04-21   Modified: 2020-03-30  

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