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Analysis of lineage-specific chromosomal conformation by in vitro enChIP

Research Project

Project/Area Number 18K06176
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43050:Genome biology-related
Research InstitutionHirosaki University

Principal Investigator

Fujita Toshitsugu  弘前大学, 医学研究科, 准教授 (10550030)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
KeywordsChIP / ChIP-Seq / locus-specific ChIP / CRISPR / enChIP / in vitro enChIP / in vitro enChIP-Seq / ゲノム機能解析 / 染色体 / 染色体三次元構造 / 遺伝子発現制御
Outline of Final Research Achievements

We previously developed in vitro enChIP, which is a biotechnology to isolate a target genomic region from cells. In this study, we combined in vitro enChIP with next-generation sequencing analysis (in vitro enChIP-Seq) to analyze B cell-specific chromosomal conformation. We focused on the Pax5 gene that encodes a master regulator of B cell-lineage commitment and succeeded in identifying genomic regions that interact commonly with the mouse and human Pax5 gene by in vitro enChIP-Seq. In addition, we analyzed histone marks on identified interacting genomic regions by ChIP-Seq and deleted those regions by genome editing to analyze physiological importance of the genomic interactions for Pax5 expression and B cell homeostasis.

Academic Significance and Societal Importance of the Research Achievements

本研究成果は、in vitro enChIP-Seq法が、特定遺伝子座を中心とした染色体三次元構造の解析に有用な技術であることを示している。in vitro enChIP-Seq法は、高度なバイオインフォマティクス処理を必要せずにゲノム領域間相互作用が解析できる技術であることから、3C法やその派生法と比較して汎用性が高い技術と考える。Pax5遺伝子座の転座が、B細胞リンパ腫に関連することが報告されていることから、本研究成果は、B細胞特異的な染色体三次元構造の生理的意義の解明のみならず、染色体三次元構造と転座の関連性の解明につながる可能性も秘めている。

Report

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

    (9 results)

All 2021 2019 Other

All Int'l Joint Research (3 results) Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (2 results) Remarks (3 results)

  • [Int'l Joint Research] the Health Institute Carlos III(スペイン)

    • Related Report
      2020 Annual Research Report
  • [Int'l Joint Research] the Health Institute Carlos III(スペイン)

    • Related Report
      2019 Research-status Report
  • [Int'l Joint Research] Lund University(スウェーデン)

    • Related Report
      2018 Research-status Report
  • [Journal Article] Locus-Specific Genomic DNA Purification Using the CRISPR System: Methods and Applications2021

    • Author(s)
      Hirotaka Fujita, Toshitsugu Fujita, and Hodaka Fujii
    • Journal Title

      The CRISPR Journal

      Volume: 4 Issue: 2 Pages: 290-300

    • DOI

      10.1089/crispr.2020.0038

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 遺伝子座特異的ChIP法によるゲノム領域間相互作用の検出2019

    • Author(s)
      藤田敏次、藤井穂高
    • Organizer
      第13回日本エピジェネティクス研究会年会
    • Related Report
      2019 Research-status Report
  • [Presentation] Identification of chromosomal interactions by locus-specific ChIP2019

    • Author(s)
      藤田敏次
    • Organizer
      第1回弘前メディカルサイエンスフォーラム
    • Related Report
      2019 Research-status Report
  • [Remarks] http://www.med.hirosaki-u.ac.jp/~bgb/

    • Related Report
      2020 Annual Research Report
  • [Remarks] http://www.med.hirosaki-u.ac.jp/~bgb/

    • Related Report
      2019 Research-status Report
  • [Remarks] http://www.med.hirosaki-u.ac.jp/~bgb/

    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2022-01-27  

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