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Establishment of a new method for functional analysis of cis-regulatory modules using CRISPRi

Research Project

Project/Area Number 26650087
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Developmental biology
Research InstitutionOkinawa Institute of Science and Technology Graduate University

Principal Investigator

Yasuoka Yuuri  沖縄科学技術大学院大学, マリンゲノミックスユニット, 研究員 (70724954)

Research Collaborator TAIRA Masanori  東京大学, 大学院理学系研究科生物科学専攻, 准教授 (60150083)
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsゲノム編集 / 転写制御 / ツメガエル / 発生 / シス制御領域 / 転写制御関連因子 / 遺伝子発現調節 / 発生ゲノム科学 / CRISPR / CRISPR-Cas9 / 遺伝子発現制御
Outline of Final Research Achievements

It is important to analyze in vivo functions of cis-regulatory modules during embryogenesis. However, quick and effective methods for such analysis have never been established. Therefore, I examined whether CRISPRi by a dead Cas9 protein (dCas9) can regulate activities of cis-regulatory modules using Xenopus embryos. Luciferase reporter assays were performed using both dCas9 mRNA and protein. The data showed that CRISPRi could not affect the reporter gene expression. This result suggests that endogenous transcriptional machineries can bump CRISPRi/dCas9 from DNA.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (16 results)

All 2017 2016 2015 2014

All Journal Article (6 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 6 results,  Open Access: 5 results,  Acknowledgement Compliant: 3 results) Presentation (10 results) (of which Int'l Joint Research: 2 results,  Invited: 2 results)

  • [Journal Article] Foxh1 Occupies cis-Regulatory Modules Prior to Dynamic Transcription Factor Interactions Controlling the Mesendoderm Gene Program.2017

    • Author(s)
      Rebekah M. Charney, Elmira Forouzmand, Jin Sun Cho, Jessica Cheung, Kitt D. Paraiso, Yuuri Yasuoka, Shuji Takahashi, Masanori Taira, Ira L. Blitz, Xiaohui Xie, and Ken W.Y. Cho.
    • Journal Title

      Developmental Cell

      Volume: 40 Issue: 6 Pages: 595-607

    • DOI

      10.1016/j.devcel.2017.02.017

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] The chordate ancestor possessed a single copy of the Brachyury gene for notochord aquisition2017

    • Author(s)
      Inoue, J., Yasuoka, Y., Takahashi, H. Satoh, N.
    • Journal Title

      Zoological Letters

      Volume: 3 Issue: 1 Pages: 4-4

    • DOI

      10.1186/s40851-017-0064-9

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Genome evolution in the allotetraploid frog Xenopus laevis2016

    • Author(s)
      Session AM, Uno Y, Kwon T, Chapman JA, Toyoda A et al.
    • Journal Title

      Nature

      Volume: 538 Issue: 7625 Pages: 336

    • DOI

      10.1038/nature19840

    • NAID

      120006240046

    • URL

      https://pure.teikyo.jp/en/publications/7cb1b606-b88e-4144-9b58-280a85c3c635

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] The Mesoderm-Forming Gene brachyury Regulates Ectoderm-Endoderm Demarcation in the Coral Acropora digitifera2016

    • Author(s)
      Yuuri Yasuoka, Chuya Shinzato, Noriyuki Satoh
    • Journal Title

      Curent Biology

      Volume: 26 Issue: 21 Pages: 2885-2892

    • DOI

      10.1016/j.cub.2016.08.011

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Occupancy of tissue-specific cis-regulatory modules by Otx2 and TLE/Groucho for embryonic head specification2014

    • Author(s)
      Yasuoka Y, Suzuki Y, Takahashi S, Someya H, Sudou N, Haramoto Y, Cho KW, Asashima M, Sugano S, Taira M.
    • Journal Title

      Nature Communications

      Volume: 5 Issue: 1 Pages: 4322-4322

    • DOI

      10.1038/ncomms5322

    • URL

      https://localhost/en/publications/cacc3f61-a004-425f-9979-221e3b6ed24a

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Evidence for participation of GCS1 in fertilization of the starlet sea anemone Nematostella vectensis: implication of a common mechanism of sperm-egg fusion in plants and animals.2014

    • Author(s)
      Ebchuqin E, Yokota N, Yamada L, Yasuoka Y, Akasaka M, Arakawa M, Deguchi R, Mori T, Sawada H.
    • Journal Title

      Biochem Biophys Res Commun

      Volume: 451 Issue: 4 Pages: 522-528

    • DOI

      10.1016/j.bbrc.2014.08.006

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 脊椎動物におけるBrachyuryの遺伝子重複がもつ 進化的意義2016

    • Author(s)
      安岡有理
    • Organizer
      第10回日本ツメガエル研究集会
    • Place of Presentation
      OISTシーサイドハウス(沖縄県国頭郡恩納村)
    • Year and Date
      2016-11-19
    • Related Report
      2016 Annual Research Report
  • [Presentation] The mesoderm-forming gene, brachyury, regulates ectoderm-endoderm demarcation in a diploblast, the coral, Acropora digitifera2016

    • Author(s)
      Yuuri Yasuoka, Chuya Shinzato, and Noriyuki Satoh
    • Organizer
      第22回国際動物学会議・第87回日本動物学会合同大会
    • Place of Presentation
      宜野湾コンベンションセンター(沖縄県宜野湾市)
    • Year and Date
      2016-11-17
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] The "mesodermal" gene brachyury regulates ecotoderm-endoderm demarcation in a diploblastic animal, the scleractinian coral Acropora digitifera2015

    • Author(s)
      Yuuri Yasuoka, Ryo Koyanagi, Yuki Yasuoka, Chuya Shinzato, Nori Satoh
    • Organizer
      Academia Sinica - OIST Joint Meeting
    • Place of Presentation
      台湾中央研究院(台湾・台北)
    • Year and Date
      2015-12-15
    • Related Report
      2015 Research-status Report
    • Int'l Joint Research
  • [Presentation] Xenopus laevis全ゲノム解析:転写因子をコードする遺伝子群の初期発生および成体器官における発現パターンの解析2015

    • Author(s)
      渡部稔、回渕修治、安岡有理、伊藤道彦、近藤真理子、越智陽樹、荻野肇、福井彰雅、平良眞規、木下勉
    • Organizer
      第38回日本分子生物学会年会
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] Xenopusにおける二つのbrachyury遺伝子の機能分化2015

    • Author(s)
      安岡有理
    • Organizer
      第1回次世代両生類研究会
    • Place of Presentation
      岡崎コンファレンスセンター
    • Year and Date
      2015-08-24
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] 脊椎動物胚における中胚葉遺伝子Brachyuryは二胚葉動物ミドリイシサンゴの胚発生において外胚葉と内胚葉の分離に働く2015

    • Author(s)
      安岡有理、小柳亮、安岡由貴、新里宙也、佐藤矩行
    • Organizer
      日本進化学会第17回大会
    • Place of Presentation
      中央大学後楽園キャンパス
    • Year and Date
      2015-08-21
    • Related Report
      2015 Research-status Report
  • [Presentation] Xenopusにおける二つのbrachyury遺伝子の機能分化から脊索におけるBrachyuryの役割を解明する2015

    • Author(s)
      Yuuri Yasuoka, Ryo Koyanagi, Nori Satoh
    • Organizer
      日本発生生物学会第48回大会
    • Place of Presentation
      つくば国際会議場
    • Year and Date
      2015-06-03
    • Related Report
      2015 Research-status Report
  • [Presentation] Transcriptional basis of the head organizer in the amphibian gastrula: ChIP seq analysis of the head organizer transcription facotors, Otx2, Lim1 and Gsc2014

    • Author(s)
      Yasuoka, Y., Suzuki, Y., Takahashi, S., Someya, H., Sudou, N., Haramoto, Y., Asashima, M., Sugano, S. and Taira, M.
    • Organizer
      第85回日本動物学会年会
    • Place of Presentation
      東北大学川内北キャンパス(宮城県仙台市)
    • Year and Date
      2014-09-12
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] Transcriptional basis of the head organizer in the amphibian gastrula: ChIP seq analysis of the head organizer transcription facotors, Otx2, Lim1 and Gsc2014

    • Author(s)
      Yasuoka, Y
    • Organizer
      15th International Xenopus Conference
    • Place of Presentation
      Pacific Grove, CA, USA
    • Year and Date
      2014-08-26
    • Related Report
      2014 Research-status Report
  • [Presentation] Functional analysis of the T-box transcription factor Brachyury in coral gastrula embryos2014

    • Author(s)
      Yasuoka, Y., Koyanagi, R., Shinzato, C. and Satoh, N.
    • Organizer
      第47回日本発生生物学会年会
    • Place of Presentation
      ウィンクあいち(愛知県名古屋市)
    • Year and Date
      2014-05-28
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2018-03-22  

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