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Development of genome-wide digital quantification method for non-coding RNAs which has single-base identification and single-molecule level quantification

Research Project

Project/Area Number 15K14425
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Genome biology
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

SHIROGUCHI KATSUYUKI  国立研究開発法人理化学研究所, 生命システム研究センター, ユニットリーダー (00454059)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsシークエンシング / デジタル計測 / バーコード / シークエンス
Outline of Final Research Achievements

Our goal was to develop a new digital RNA sequencing which could be applied for non-polyA tailed RNA and had single base resolution. Then, we aim to make a standardized high throughput digital RNA sequencing for researcher all over the world.
In this study, we attached one of many different molecular barcodes to each target molecules stochastically, amplified these molecules, and sequenced them. By counting the number of different barcodes, we quantified the original (i.e., before amplification) number of target molecules. Since the sequenced target molecules that have the same barcode sequence are from the same original molecules, we identified errors, and finally, we determined target sequences with single base resolution. I think this method will be used for quantification of nucleic acid molecules in many measurements.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • Research Products

    (8 results)

All 2017 2016 2015

All Journal Article (2 results) (of which Peer Reviewed: 1 results) Presentation (5 results) (of which Int'l Joint Research: 2 results,  Invited: 3 results) Book (1 results)

  • [Journal Article] デジタルRNAシークエンシング ゲノムワイドな1分子定量法2016

    • Author(s)
      城口克之
    • Journal Title

      生物と化学(日本農芸化学会)

      Volume: 54 Pages: 787-788

    • NAID

      130006176426

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 次世代シークエンサーを用いたDNA, RNAの網羅的デジタル計測2016

    • Author(s)
      城口克之
    • Journal Title

      現代化学

      Volume: 546 Pages: 47-50

    • Related Report
      2016 Annual Research Report
  • [Presentation] 1細胞レベル・塩基分解能でのハイスープット菌叢解析2017

    • Author(s)
      城口克之、Jianshi Jin
    • Organizer
      第11回日本ゲノム微生物学会年会
    • Place of Presentation
      慶應義塾大学(神奈川県、藤沢市)
    • Year and Date
      2017-03-02
    • Related Report
      2016 Annual Research Report
  • [Presentation] Finding genomic minority cells by sequencing2016

    • Author(s)
      城口克之
    • Organizer
      第54回日本生物物理学会年会
    • Place of Presentation
      つくば国際会議場(茨城県、つくば市)
    • Year and Date
      2016-11-25
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] High Throughput Single Cell Analysis for Target Genomic Sequences2016

    • Author(s)
      城口克之
    • Organizer
      International conference on Single Cell Research 2016
    • Place of Presentation
      東京大学(東京都、文京区)
    • Year and Date
      2016-11-16
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] A Novel Method for High Throughput Single Cell Identification and Counting by Target Gene Sequencing2016

    • Author(s)
      城口克之
    • Organizer
      QBiC Symposium 2016
    • Place of Presentation
      千里ライフサイエンスセンター(大阪府、豊中市)
    • Year and Date
      2016-09-06
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Genome-wide Digital mRNA Counting Using "Molecular Barcoding" and Its Application for Differentiation Study2015

    • Author(s)
      Katsuyuki Shiroguchi
    • Organizer
      BIT’s 5th Annual World Congress of Molecular & Cell Biology -2015
    • Place of Presentation
      Nangjing (China)
    • Year and Date
      2015-04-26
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research / Invited
  • [Book] JSI Newsletter2016

    • Author(s)
      城口克之
    • Total Pages
      1
    • Publisher
      日本免疫学会
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2018-03-22  

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