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Development of a non-destructive method to track the transcriptome of living cells across time

Research Project

Project/Area Number 22K19275
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionKyushu University

Principal Investigator

Yasuyuki Ohkawa  九州大学, 生体防御医学研究所, 教授 (80448430)

Project Period (FY) 2022-06-30 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2023: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2022: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Keywordsトランスクリプトーム / ライブセル / 系譜追跡技術
Outline of Research at the Start

細胞は、エクソソームと呼ばれる細胞外分泌小胞を産生する。特に、エクソソームは細胞質の様々な物質を含んでおり微量にRNAを含有していることが知られているが、その解明はほとんど進んでいない。我々は先行知見を基に、細胞外のエクソソームに含まれる極微量のRNAを安定的に解析する手法を確立することで生細胞の非破壊的RNAseq法を開発する。培養細胞レベルで細胞集団の解析をする手法を開発し、更に個々の細胞をバーコード技術で識別し、分泌した個々のエクソソームを単一エクソソームRNA解析することで、単一細胞レベルのライブセルRNAseq法の開発を行う。

Outline of Final Research Achievements

The phenotype of an individual is established through development and differentiation and changes dynamically until it decays, leading to aging and death. Transcriptome analysis has been widely used as the information that best reflects the state of the changing phenotype. On the other hand, current transcriptome analysis cannot follow changes in the same cell because it requires the destruction of the cell and extraction of RNA. In this study, we developed a non-destructive RNAseq method for living cells based on our own previous findings that endogenous RNA correlates with a small amount of RNA contained in exosomes secreted by cells. The project has been successfully completed and is now being applied to a variety of samples.

Academic Significance and Societal Importance of the Research Achievements

本研究が基盤とする技術は微量RNA操作技術である。現在単一細胞トランスクリプトームが注目を浴びる中、その多くはキット化された商用プラットフォームに依存しており汎用的な反面、微量RNAを操作する場面を研究者から奪っている一面が存在する。本研究が行うごく少量のRNAを扱う基礎技術のプロトコール化と普及は次の新たな解析技術の基盤となる基礎研究能力の喪失を防ぎ、今後のニーズに即した研究基盤の強化につながると考えている。

Report

(3 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Research-status Report
  • Research Products

    (47 results)

All 2024 2023 2022 Other

All Journal Article (19 results) (of which Int'l Joint Research: 5 results,  Peer Reviewed: 19 results,  Open Access: 19 results) Presentation (24 results) (of which Int'l Joint Research: 6 results,  Invited: 23 results) Book (2 results) Remarks (2 results)

  • [Journal Article] Production of a Monoclonal Antibody for Histone H2b Isoform H2b3b2024

    • Author(s)
      Egashira Saki、Tachibana Taro、Nakamura Mako、Ohkawa Yasuyuki、Harada Akihito
    • Journal Title

      Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

      Volume: 43 Issue: 2 Pages: 75-80

    • DOI

      10.1089/mab.2023.0025

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genome-wide ATAC-see screening identifies TFDP1 as a modulator of global chromatin accessibility2024

    • Author(s)
      Ishii Satoko、Kakizuka Taishi、Park Sung-Joon、Tagawa Ayako、Sanbo Chiaki、Tanabe Hideyuki、Ohkawa Yasuyuki、Nakanishi Mahito、Nakai Kenta、Miyanari Yusuke
    • Journal Title

      Nature Genetics

      Volume: 56 Issue: 3 Pages: 473-482

    • DOI

      10.1038/s41588-024-01658-1

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genome-wide mapping and cryo-EM structural analyses of the overlapping tri-nucleosome composed of hexasome-hexasome-octasome moieties2024

    • Author(s)
      Nishimura Masahiro、Fujii Takeru、Tanaka Hiroki、Maehara Kazumitsu、Morishima Ken、Shimizu Masahiro、Kobayashi Yuki、Nozawa Kayo、Takizawa Yoshimasa、Sugiyama Masaaki、Ohkawa Yasuyuki、Kurumizaka Hitoshi
    • Journal Title

      Communications Biology

      Volume: 7 Issue: 1 Pages: 61-61

    • DOI

      10.1038/s42003-023-05694-1

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Precise immunofluorescence canceling for highly multiplexed imaging capturing specific cell state2023

    • Author(s)
      Tomimatsu Kosuke、Fujii Takeru、Bise Ryoma、Hosoda Kazufumi、Taniguchi Yosuke、Ochiai Hiroshi、Ohishi Hiroaki、Ando Kanta、Minami Ryoma、Tanaka Kaori、Tachibana Taro、Mori Seiichi、Harada Akihito、Maehara Kazumitsu、Nagasaki Masao、Uchida Seiichi、Kimura Hiroshi、Narita Masashi、Ohkawa Yasuyuki
    • Journal Title

      bioRxiv

      Volume: -

    • DOI

      10.1101/2023.10.17.561810

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Cryo-EM and biochemical analyses of the nucleosome containing the human histone H3 variant H3.82023

    • Author(s)
      Hirai Seiya、Kujirai Tomoya、Akatsu Munetaka、Ogasawara Mitsuo、Ehara Haruhiko、Sekine Shun-ichi、Ohkawa Yasuyuki、Takizawa Yoshimasa、Kurumizaka Hitoshi
    • Journal Title

      The Journal of Biochemistry

      Volume: 174 Issue: 6 Pages: 549-559

    • DOI

      10.1093/jb/mvad069

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Phase-separated nuclear bodies of nucleoporin fusions promote condensation of MLL1/CRM1 and rearrangement of 3D genome structure2023

    • Author(s)
      Oka Masahiro、Otani Mayumi、Miyamoto Yoichi、Oshima Rieko、Adachi Jun、Tomonaga Takeshi、Asally Munehiro、Nagaoka Yuya、Tanaka Kaori、Toyoda Atsushi、Ichikawa Kazuki、Morishita Shinichi、Isono Kyoichi、Koseki Haruhiko、Nakato Ryuichiro、Ohkawa Yasuyuki、Yoneda Yoshihiro
    • Journal Title

      Cell Reports

      Volume: 42 Issue: 8 Pages: 112884-112884

    • DOI

      10.1016/j.celrep.2023.112884

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] FOXK1 promotes nonalcoholic fatty liver disease by mediating mTORC1-dependent inhibition of hepatic fatty acid oxidation.2023

    • Author(s)
      Fujinuma S, Nakatsumi H, Shimizu H, Sugiyama S, Harada A, Goya T, Tanaka M, Kohjima M, Takahashi M, Izumi Y, Yagi M, Kang D, Kaneko M, Shigeta M, Bamba T, Ohkawa Y, Nakayama KI*.
    • Journal Title

      Cell Reports

      Volume: 42(5) Issue: 5 Pages: 112530-112530

    • DOI

      10.1016/j.celrep.2023.112530

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Design and implementation of a hybrid cloud system for large-scale human genomic research.2023

    • Author(s)
      Masao Nagasaki, Yayoi Sekiya, Akihiro Asakura, Ryo Teraoka, Ryoko Otokozawa, Hiroki Hashimoto, Takahisa Kawaguchi, Keiichiro Fukazawa, Yuichi Inadomi, Ken T Murata, Yasuyuki Ohkawa, Izumi Yamaguchi, Takamichi Mizuhara, Katsushi Tokunaga, Yuji Sekiya, Toshihiro Hanawa, Ryo Yamada, Fumihiko Matsuda.
    • Journal Title

      Hum Genome Var.

      Volume: 10 Issue: 1 Pages: 6-6

    • DOI

      10.1038/s41439-023-00231-2

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Senescence atlas reveals an aged-like inflamed niche that blunts muscle regeneration.2023

    • Author(s)
      Moiseeva V, Cisneros A, Sica V, Deryagin O, Lai Y, Jung S, Andres E, An J, Segales J, Ortet L, Lukesova V, Volpe G, Benguria A, Dopazo A, Benitah SA, Urano Y, Del Sol A, Esteban MA, Ohkawa Y, Serrano AL, Perdiguero E, Munoz-Canoves P.
    • Journal Title

      Nature

      Volume: 613 Issue: 7942 Pages: 169-178

    • DOI

      10.1038/s41586-022-05535-x

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] STREAMING-tag system reveals spatiotemporal relationships between transcriptional regulatory factors and transcriptional activity2022

    • Author(s)
      Ohishi Hiroaki、Shimada Seiru、Uchino Satoshi、Li Jieru、Sato Yuko、Shintani Manabu、Owada Hitoshi、Ohkawa Yasuyuki、Pertsinidis Alexandros、Yamamoto Takashi、Kimura Hiroshi、Ochiai Hiroshi
    • Journal Title

      Nature Communications

      Volume: 13 Issue: 1 Pages: 7672-7672

    • DOI

      10.1038/s41467-022-35286-2

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Prostaglandin E2 receptor Ptger4b regulates female-specific peptidergic neurons and female sexual receptivity in medaka2022

    • Author(s)
      Fleming Thomas、Kikuchi Yukiko、Nakajo Mikoto、Tachizawa Masaya、Inazumi Tomoaki、Tsuchiya Soken、Sugimoto Yukihiko、Saito Daisuke、Suyama Mikita、Ohkawa Yasuyuki、Baba Takashi、Morohashi Ken-ichirou、Okubo Kataaki
    • Journal Title

      Communications Biology

      Volume: 5 Issue: 1 Pages: 1215-1215

    • DOI

      10.1038/s42003-022-04195-x

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Drosophila transcription factor NF-Y suppresses transcription of the lipase 4 gene, a key gene for lipid storage.2022

    • Author(s)
      Yasuhide Yoshioka, Keisuke Anzai, Ryosuke Kowada, Ken Hiratsuka, Teppei Hirayabu, Masashi Yasuda, Yasuyuki Ohkawa, Tetsuya Sato, Mikita Suyama, Hideki Yoshida, Masamitsu Yamaguchi.
    • Journal Title

      Exp Cell Res.

      Volume: 420 Issue: 1 Pages: 113307-113307

    • DOI

      10.1016/j.yexcr.2022.113307

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Tmsb10 triggers fetal Leydig differentiation by suppressing the RAS/ERK pathway.2022

    • Author(s)
      Miki Inoue, Takashi Baba, Fumiya Takahashi, Miho Terao, Shogo Yanai, Yuichi Shima, Daisuke Saito, Kei Sugihara, Takashi Miura, Shuji Takada, Mikita Suyama, Yasuyuki Ohkawa, Ken-Ichirou Morohashi.
    • Journal Title

      Commun Biol.

      Volume: 5 Issue: 1 Pages: 974-974

    • DOI

      10.1038/s42003-022-03941-5

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Identification of a drug-response gene in multiple myeloma through longitudinal single-cell transcriptome sequencing2022

    • Author(s)
      Masuda Toru、Haji Shojiro、Nakashima Yasuhiro、Tsuda Mariko、Kimura Daisaku、Takamatsu Akiko、Iwahashi Norifusa、Umakoshi Hironobu、Shiratsuchi Motoaki、Kikutake Chie、Suyama Mikita、Ohkawa Yasuyuki、Ogawa Yoshihiro
    • Journal Title

      iScience

      Volume: 25 Issue: 8 Pages: 104781-104781

    • DOI

      10.1016/j.isci.2022.104781

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Uterus-specific transcriptional regulation underlies eggshell pigment production in Japanese quail2022

    • Author(s)
      Ishishita Satoshi、Kitahara Shumpei、Takahashi Mayuko、Iwasaki Sakura、Tatsumoto Shoji、Hara Izumi、Kaneko Yoshiki、Kinoshita Keiji、Yamaguchi Katsushi、Harada Akihito、Ohmori Yasushige、Ohkawa Yasuyuki、Go Yasuhiro、Shigenobu Shuji、Matsuda Yoichi、Suzuki Takayuki
    • Journal Title

      PLOS ONE

      Volume: 17 Issue: 3 Pages: 0265008-0265008

    • DOI

      10.1371/journal.pone.0265008

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Photo-isolation chemistry for high-resolution and deep spatial transcriptome with mouse tissue sections2022

    • Author(s)
      Honda Mizuki、Kimura Ryuichi、Harada Akihito、Maehara Kazumitsu、Tanaka Kaori、Ohkawa Yasuyuki、Oki Shinya
    • Journal Title

      STAR Protocols

      Volume: 3 Issue: 2 Pages: 101346-101346

    • DOI

      10.1016/j.xpro.2022.101346

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A spinal microglia population involved in remitting and relapsing neuropathic pain2022

    • Author(s)
      Kohno Keita、Shirasaka Ryoji、Yoshihara Kohei、Mikuriya Satsuki、Tanaka Kaori、Takanami Keiko、Inoue Kazuhide、Sakamoto Hirotaka、Ohkawa Yasuyuki、Masuda Takahiro、Tsuda Makoto
    • Journal Title

      Science

      Volume: 376 Issue: 6588 Pages: 86-90

    • DOI

      10.1126/science.abf6805

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Tenogenic Induction From Induced Pluripotent Stem Cells Unveils the Trajectory Towards Tenocyte Differentiation2022

    • Author(s)
      Yoshimoto Yuki、Uezumi Akiyoshi、Ikemoto-Uezumi Madoka、Tanaka Kaori、Yu Xinyi、Kurosawa Tamaki、Yambe Shinsei、Maehara Kazumitsu、Ohkawa Yasuyuki、Sotomaru Yusuke、Shukunami Chisa
    • Journal Title

      Frontiers in Cell and Developmental Biology

      Volume: 10 Pages: 780038-780038

    • DOI

      10.3389/fcell.2022.780038

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Direct Conversion of Human Endothelial Cells Into Liver Cancer-Forming Cells Using Nonintegrative Episomal Vectors.2022

    • Author(s)
      Goya T, Horisawa K, Udono M, Ohkawa Y, Ogawa Y, Sekiya S, Suzuki A.
    • Journal Title

      Hepatol Commun.

      Volume: - Issue: 7 Pages: 1725-1740

    • DOI

      10.1002/hep4.1911

    • Related Report
      2022 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] 連続免疫染色法による高解像度空間オミクスの開発2024

    • Author(s)
      大川恭行
    • Organizer
      ゲノム創薬・創発フォーラム
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] 組織形成を理解するための単一細胞マルチオミクスの開発2024

    • Author(s)
      大川恭行
    • Organizer
      創薬医学講座
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] 組織形成を理解するための単一細胞マルチオミクスの開発2023

    • Author(s)
      原田 哲仁、富松航佑、藤井健、大川恭行
    • Organizer
      第46回日本分子生物学会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] Developing single-cell multi-omics to understand tissue formation2023

    • Author(s)
      Yasuyuki Ohkawa
    • Organizer
      サルコペニア研究会
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Comprehensive Analysis of Signal Transduction Activation Pathways through Sequential Immunostaining2023

    • Author(s)
      Yasuyuki Ohkawa
    • Organizer
      Single Cell Genomics Conference 2023
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 遺伝子発現制御を包括的に解析する技術の開発2023

    • Author(s)
      大川恭行
    • Organizer
      2023年度国立遺伝学研究所クロマチン研究会(NIG-JOINT)
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] 組織形成を理解するための単一細胞マルチオミクスの開発2023

    • Author(s)
      大川恭行
    • Organizer
      奈良県立医科大学講演
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] トランスクリプトミクスによる筋再生過程の高深度解析2023

    • Author(s)
      大川恭行
    • Organizer
      第9回日本筋学会学術集会/第10回筋ジストロフィー医療研究会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] 組織形成を理解するための単一細胞マルチオミクスの開発2023

    • Author(s)
      大川恭行
    • Organizer
      生物科学セミナー
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] Development of spatial multi-omics for understanding skeletal muscle aging2023

    • Author(s)
      Yasuyuki Ohkawa
    • Organizer
      1st Symposium on "Skeletal muscle cells in Growth and Disease"
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 単一細胞マルチオミクスによる骨格筋分化制御機構の解明2023

    • Author(s)
      大川恭行
    • Organizer
      第6回運動器と健康研究会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] Spatial multi-omics for understanding chromatin dynamics2023

    • Author(s)
      大川恭行
    • Organizer
      2023年度高深度オミクスシンポジウム
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] データから変化を取り出す新たな単一細胞解析技術2023

    • Author(s)
      大川恭行
    • Organizer
      【富士フイルム】講演
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] Single-cell multi-targeted chromatin integration labeling technology for understanding chromatin dynamics2022

    • Author(s)
      Yasuyuki Ohkawa
    • Organizer
      CSHA meeting on Integrative Epigenetics in Plants
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 空間オミクス技術によるクロマチン構造制御メカニズムの解明2022

    • Author(s)
      大川恭行
    • Organizer
      第95回日本生化学会大会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] 細胞恒常性を担うシグナルと遺伝子発現の制御2022

    • Author(s)
      大川恭行
    • Organizer
      第16回日本臨床ストレス応答学会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] 高深度解析による全ゲノムレベルの単一細胞オミクスへの挑戦2022

    • Author(s)
      大川恭行
    • Organizer
      NGS EXPO 2022
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] 単一細胞軌跡追跡によるクロマチンダイナミクスの解明2022

    • Author(s)
      大川恭行
    • Organizer
      2022年度国立遺伝学研究所クロマチン研究会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] Chromatin regulation during skeletal muscle regeneration2022

    • Author(s)
      Yasuyuki Ohkawa
    • Organizer
      第17回生命医科学研究所ネットワーク国際シンポジウム
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 骨格筋分化のクロマチンダイナミクス2022

    • Author(s)
      大川恭行
    • Organizer
      日本遺伝学会第94回大会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] トランスクリプトミクスで迫る細胞分化能2022

    • Author(s)
      大川恭行
    • Organizer
      大阪大学産業科学研究所セミナー
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] Spatial multi-omics for understanding gene expression regulated by cell-cell interaction.2022

    • Author(s)
      Yasuyuki Ohkawa
    • Organizer
      Japan-UK Regulation through Chromatin Conference
    • Related Report
      2022 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 空間オミクスによる骨格筋組織老化機序の解明2022

    • Author(s)
      大川恭行
    • Organizer
      第8回日本筋学会学術集会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Presentation] クロマチン解析のための新技術開発の現状と今後の展開2022

    • Author(s)
      大川恭行
    • Organizer
      第15回日本エピジェネティクス研究会年会
    • Related Report
      2022 Research-status Report
    • Invited
  • [Book] 月刊細胞2023

    • Author(s)
      原田 哲仁,大川 恭行
    • Total Pages
      4
    • Publisher
      ニューサイエンス社
    • Related Report
      2023 Annual Research Report
  • [Book] 生体の科学2023

    • Author(s)
      原田 哲仁, 富松 航佑, 武 千メイ, 大川 恭行
    • Total Pages
      5
    • Publisher
      公益財団法人金原一郎記念医学医療振興財団・医学書院
    • Related Report
      2023 Annual Research Report
  • [Remarks] 九州大学 トランスクリプトミクス分野ホームページ

    • URL

      https://tx.bioreg.kyushu-u.ac.jp/

    • Related Report
      2023 Annual Research Report
  • [Remarks] 九州大学 生体防御医学研究所 トランスクリプトミクス分野HP

    • URL

      https://tx.bioreg.kyushu-u.ac.jp/

    • Related Report
      2022 Research-status Report

URL: 

Published: 2022-07-05   Modified: 2025-01-30  

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