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Molecular analysis and universality exploration of DNA damage-triggered reprogramming to stem cells

Research Project

Project/Area Number 21F21084
Research Category

Grant-in-Aid for JSPS Fellows

Allocation TypeSingle-year Grants
Section外国
Review Section Basic Section 44030:Plant molecular biology and physiology-related
Research InstitutionUtsunomiya University

Principal Investigator

玉田 洋介  宇都宮大学, 工学部, 准教授 (50579290)

Co-Investigator(Kenkyū-buntansha) GU NAN  宇都宮大学, 工学部, 外国人特別研究員
Project Period (FY) 2021-04-28 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 2022: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2021: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsStem cell / Reprogramming / Physcomitrium patens / DNA damage / Single-nucleus RNA-seq / snRNA-sequencing
Outline of Research at the Start

We found that transient DNA damage can induce the reprogramming of differentiated leaf cells into stem cells without wounding and exogenous phytohormones in the moss Physcomitrium patens. This depends on the DNA damage sensor kinase ATR and the reprogramming signal integrators STEMINs. In this project, we will investigate the molecular mechanism of how DNA damage induces STEMIN1 through ATR only in a part of leaf cells. As DNA damage response is conserved in plants and animals, we will also examine whether DNA damage can enhance the reprogramming in other organisms.

Outline of Annual Research Achievements

Our study aimed to understand the molecular mechanism of DNA damage-triggered reprogramming from differentiated leaf cells into stem cells in Physcomitrium patens. To this end, we conducted single cell transcriptome analysis through single nucleus RNA-sequencing (snRNA-seq) to track transcriptional changes during the reprogramming process. We successfully obtained high quality snRNA-seq data. After the clustering of nuclear transcriptomes, we obtained a set of genes which were specifically expressed in each cluster. With fluorescence observation of promoter reporter lines of these specifically expressed genes, multiple clusters were annotated to specific tissues. Based on these cluster annotation results, we can extract the reprogramming trajectory of nuclei from leaf cells to newly formed protonema stem cells. From this analysis, we can identify stage-specific genes during DNA damage-triggered reprogramming and differentially expressed genes between reprogramming and non-reprogramming leaf cells at the late stage, which will provide a potential list of key genes driving the cellular reprogramming.
To explore the universality of DNA damage-triggered reprogramming in land plants, we established culture conditions for multiple angiosperms and treated each plant with Zeocin. In most cases, the DNA damage reagent caused cell death in the tested plants, suggesting that efficient DNA damage repair is critical for successful cellular reprogramming.

Research Progress Status

令和4年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

令和4年度が最終年度であるため、記入しない。

Report

(2 results)
  • 2022 Annual Research Report
  • 2021 Annual Research Report
  • Research Products

    (14 results)

All 2022 2021 Other

All Int'l Joint Research (1 results) Journal Article (1 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 1 results,  Open Access: 1 results) Presentation (6 results) (of which Int'l Joint Research: 4 results,  Invited: 1 results) Remarks (6 results)

  • [Int'l Joint Research] Huazhong Agricultural University(中国)

    • Related Report
      2021 Annual Research Report
  • [Journal Article] Topoisomerase 1α is required for synchronous spermatogenesis in Physcomitrium patens2022

    • Author(s)
      Nan Gu、Chunli Chen、Yukiko Kabeya、Mitsuyasu Hasebe、Yosuke Tamada
    • Journal Title

      New Phytologist

      Volume: 234 Issue: 1 Pages: 137-148

    • DOI

      10.1111/nph.17983

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] DNA damage triggers stem cell formation in the moss Physcomitrium patens2022

    • Author(s)
      Nan Gu, Gergo Palfalvi, Yosuke Tamada
    • Organizer
      Integrative Epigenetics in Plants (Cold Spring Harbor Asia)
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 3D image analysis of spermatid nuclei to understand the mechanism of the sperm formation in a land plant2022

    • Author(s)
      Yosuke Tamada, Nan Gu
    • Organizer
      Biomedical Imaging and Sensing Conference 2022 (BISC2022)
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 顕微鏡イメージング・細胞操作による植物の高い再生能力の解明2022

    • Author(s)
      玉田洋介, Nan Gu
    • Organizer
      宇都宮大学第3回コラボレーションフェア
    • Related Report
      2022 Annual Research Report
  • [Presentation] Topoisomerase 1α is required for synchronous spermatogenesis in Physcomitrium patens2021

    • Author(s)
      Nan Gu, Chunli Chen, Yukiko Kabeya, Mitsuyasu Hasebe, Yosuke Tamada
    • Organizer
      2021中国植物生物学大会
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Fluorescence imaging to understand the molecular mechanism of DNA damage-triggered cellular reprogramming in plants2021

    • Author(s)
      Yosuke Tamada, Akihiro Imai, Nan Gu
    • Organizer
      Biomedical Imaging and Sensing Conference 2021 (BISC2021)
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Reprogramming of differentiated cells to stem cells triggered by DNA damage2021

    • Author(s)
      Yosuke Tamada, Akihiro Imai, Nan Gu
    • Organizer
      The 44th Annual Meeting of the Molecular Biology Society of Japan
    • Related Report
      2021 Annual Research Report
  • [Remarks] Science Japan Article

    • URL

      https://sj.jst.go.jp/news/202206/n0601-02k.html

    • Related Report
      2022 Annual Research Report
  • [Remarks] 宇都宮大学 地域創生科学研究科 光工学プログラム

    • URL

      https://www.utsunomiya-u.ac.jp/grdc/agricultural/light_engineering.html

    • Related Report
      2021 Annual Research Report
  • [Remarks] 宇都宮大学 工学部 物質環境化学コース

    • URL

      http://www.eng.utsunomiya-u.ac.jp/course/material-and-environmental-chemistry-course/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 宇都宮大学 オプティクス教育研究センター (CORE)

    • URL

      https://uu-core.com/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 宇都宮大学 ロボティクス・工農技術研究所 (REAL)

    • URL

      https://ja-jp.facebook.com/REAL.Utsunomiya/

    • Related Report
      2021 Annual Research Report
  • [Remarks] 日本光学会 研究グループ

    • URL

      http://myosj.or.jp/group/

    • Related Report
      2021 Annual Research Report

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Published: 2021-05-27   Modified: 2024-03-26  

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