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Construction of improved virus-susceptible host cells by genome editing

Research Project

Project/Area Number 17H04003
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Environmental and hygienic pharmacy
Research InstitutionNational Institute of Infectious Diseases

Principal Investigator

Hanada Kentaro  国立感染症研究所, 品質保証・管理部, 主任研究官 (30192701)

Co-Investigator(Kenkyū-buntansha) 桶本 優子 (中村優子)  国立感染症研究所, 細胞化学部, 主任研究官 (30392319)
齊藤 恭子  国立感染症研究所, 細胞化学部, 主任研究官 (70235034)
佐久間 智理  国立感染症研究所, 細胞化学部, 研究員 (80782888)
Project Period (FY) 2017-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2020: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2019: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2018: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2017: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Keywords宿主細胞 / Vero細胞 / フラビウイルス / ポリオウイルス / 内在性レトロウイルス / レギュラトリーサイエンス / プリオン / ゲノム編集 / 感染症 / Huh7細胞 / ヒト神経芽腫由来細胞 / ワクチン / 細胞 / ゲノム / ウイルス
Outline of Final Research Achievements

The Vero cell lineage, which is universally used for infectious disease control, has multiple sub-lines. Comparative genomic analysis revealed that the cell sub-lines can be broadly classified into two lineages, but the endogenous retroviral insertion position and homozygous deletion position of the type I interferon gene cluster are identical in all four Vero cell lines examined. We generated a Vero cell line with persistent replication of the yellow fever virus 17D replicon and another Vero cell line without poliovirus growth potential, which may be useful for the WHO-led polio eradication program. We found the virological utility of Huh7.5.1-8 cells, a subline of the HuH-7 cell line derived from human hepatocellular carcinoma with high hepatitis C virus productivity, and also clarified the characteristics of RIG-I mutations in the HuH-7 cell line.

Academic Significance and Societal Importance of the Research Achievements

本研究課題の成果は、ウイルス感染症ワクチンを生産する細胞基材の開発・改良のための科学基盤となる。例えば、GFP発現型の黄熱ウイルスレプリコン持続複製Vero細胞は、同ウイルス複製を阻害または増強する薬剤や宿主細胞変異株を簡便に探索するツールとなる。また、本研究で作出したポリオウイルス増殖能のないVero細胞は、WHOが主導するポリオ根絶計画に対処しつつも、広範な種類の新興再興ウイルス感染症の原因ウイルスを分離する際の標準的な宿主細胞となりえる。さらに、Vero細胞やHuH-7細胞のゲノム解析情報はこれら細胞の品質管理に活用できる。

Report

(5 results)
  • 2021 Final Research Report ( PDF )
  • 2020 Annual Research Report
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (23 results)

All 2021 2020 2019 2018 2017 Other

All Journal Article (7 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 7 results,  Open Access: 6 results) Presentation (11 results) (of which Int'l Joint Research: 2 results,  Invited: 4 results) Remarks (5 results)

  • [Journal Article] Poliovirus-nonsusceptible Vero cell line for the World Health Organization global action plan2021

    • Author(s)
      Okemoto-Nakamura Y, Someya K, Yamaji T, Saito K, Takeda M, Hanada K
    • Journal Title

      Sci Rep

      Volume: 11 Issue: 1 Pages: 6746-6746

    • DOI

      10.1038/s41598-021-86050-3

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Seroprevalence of Flavivirus Neutralizing Antibodies in Thailand by High-Throughput Neutralization Assay: Endemic Circulation of Zika Virus before 20122021

    • Author(s)
      Yamanaka Atsushi, Matsuda Mami, Okabayashi Tamaki, Pitaksajjakul Pannamthip, Ramasoota Pongrama, Saito Kyoko, Fukasawa Masayoshi, Hanada Kentaro, Matsuura Tomokazu, Muramatsu Masamichi, Shioda Tatsuo, Suzuki Ryosuke
    • Journal Title

      mSphere

      Volume: 6 Issue: 4

    • DOI

      10.1128/msphere.00339-21

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Comparative characterization of flavivirus production in two cell lines: Human hepatoma-derived Huh7.5.1-8 and African green monkey kidney-derived Vero2020

    • Author(s)
      Kyoko Saito, Masayoshi Fukasawa, Yoshitaka Shirasago, Ryosuke Suzuki, Naoki Osada, Toshiyuki Yamaji, Takaji Wakita, Eiji Konishi, Kentaro Hanada
    • Journal Title

      PLOS ONE

      Volume: 15 Issue: 4 Pages: e0232274-e0232274

    • DOI

      10.1371/journal.pone.0232274

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Identification of Characteristic Genomic Markers in Human Hepatoma HuH-7 and Huh7.5.1-8 Cell Lines2020

    • Author(s)
      Kawamoto M#, Yamaji T#, Saito K, Shirasago Y, Satomura K, Endo T, Fukasawa M, Hanada K, Osada N.
    • Journal Title

      Front Genet

      Volume: 11 Pages: 546106-546106

    • DOI

      10.3389/fgene.2020.546106

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 1.A PRNP-Disrupted Human Neuroblastoma Cell Line and Its Stable Transformants Expressing Prion Protein Variants2019

    • Author(s)
      Yuko Okemoto-Nakamura , Isei Tanida, Toshiyuki Yamaji , Kentaro Hanada , Ken’ichi Hagiwara
    • Journal Title

      BPB Reports

      Volume: 2 Pages: 73-79

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Thermostable hepatitis C virus JFH1-derived variant isolated by adaptation to Huh7.5.1 cells2018

    • Author(s)
      Shirasago Yoshitaka、Fukazawa Hidesuke、Aizaki Hideki、Suzuki Tetsuro、Suzuki Takeru、Sugiyama Kazuo、Wakita Takaji、Hanada Kentaro、Abe Ryo、Fukasawa Masayoshi
    • Journal Title

      Journal of General Virology

      Volume: 99 Issue: 10 Pages: 1407-1417

    • DOI

      10.1099/jgv.0.001117

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Novel endogenous simian retroviral integrations in Vero cells: implications for quality control of a human vaccine cell substrate2018

    • Author(s)
      Sakuma Chisato、Sekizuka Tsuyoshi、Kuroda Makoto、Kasai Fumio、Saito Kyoko、Ikeda Masaki、Yamaji Toshiyuki、Osada Naoki、Hanada Kentaro
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 644-644

    • DOI

      10.1038/s41598-017-18934-2

    • NAID

      120006401475

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 初心者のための入門講座:感染症研究へのゲノム編集技術の応用 CRISPR/Cas9システムによる新規の細胞株の創出2020

    • Author(s)
      中村(桶本)優子、 染谷健二、 齊藤恭子、 山地俊之、 竹田誠、 花田賢太郎
    • Organizer
      第69回日本感染症学会東日本地方会学術集会・第69回日本化学療法学会東日本支部総会合同学会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] バイオロジクスにおけるウイルス安全性確保の新潮流2020

    • Author(s)
      花田賢太郎
    • Organizer
      第141回日本薬学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Vero細胞ゲノムに存在するサル内在性レトロウイルスの性状解析2020

    • Author(s)
      齊藤恭子
    • Organizer
      第141回日本薬学会
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] 寄生細菌クラミジアは宿主細胞のセラミドをいかにして利用するのか2020

    • Author(s)
      花田賢太郎
    • Organizer
      日本薬学会生物系薬学部会主催 第21回Pharmaco-Hematologyシンポジウム~薬学の基礎と臨床~
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] ヒト肝癌由来Huh7.5.1-8細胞とアフリカミドリザル腎由来Vero細胞におけるフラビウイルス産生能の比較解析2019

    • Author(s)
      齊藤恭子, 深澤征義, 白砂圭崇, 鈴木亮介, 山地俊之, 脇田隆字, 小西英二, 花田賢太郎
    • Organizer
      第92回日本生化学会大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Construction of a yellow fever virus subgenomic replicon system and characterization of cell lines harboring the replicon2019

    • Author(s)
      Kyoko Saito, Masayoshi Fukasawa, Ryosuke Suzuki, Tomohiko Takasaki, Kentaro Hanada
    • Organizer
      第67回日本ウイルス学会学術集会
    • Related Report
      2019 Annual Research Report
  • [Presentation] ヒト肝癌由来Huh7.5.1-8細胞とアフリカミドリザル腎由来Vero細胞におけるフラビウイルス産生の比較解析2019

    • Author(s)
      齊藤恭子、深澤征義、白砂圭崇、鈴木亮介、脇田隆字、小西英二、花田賢太郎
    • Organizer
      日本薬学会第139年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 旧世界ザルゲノムに存在する内在性サルレトロウィルス(SERV)配列の探索と分子系統解析2018

    • Author(s)
      池田昌輝, 里村和浩, 関塚剛史, 花田賢太郎, 遠藤俊徳, 長田直樹
    • Organizer
      第90回日本遺伝学会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Search for the factors related to HCV replication in the HuH-7 cell line lineages2018

    • Author(s)
      Kawamoto M, Endo T, Fukasawa M, Hanada K, Osada N
    • Organizer
      SMBE2018
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Endogenous Simian Retrovirus Variations in Vero Cells: Implications for Quality Control of a Human Vaccine Cell Substrate2018

    • Author(s)
      Chisato Sakuma, Tsuyoshi Sekizuka, Makoto Kuroda, Fumio Kasai, Kyoko Saito, Masaki Ikeda, Toshiyuki Yamaji, Naoki Osada, Kentaro Hanada
    • Organizer
      4th International Symposium for Medicinal Sciences, 2018.3.25-28, Kanazawa
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Comparative characterization of human hepatoma-derived Huh7.5.1-8 cells and African green monkey kidney-derived Vero cells in Japanese encephalitis virus production ヒト肝癌由来Huh7.5.1-8細胞とアフリカミドリザル腎由来Vero細胞における日本脳炎ウイルス産生の比較解析2017

    • Author(s)
      齊藤恭子、深澤征義、白砂圭崇、鈴木亮介、脇田隆字、小西英二、花田賢太郎
    • Organizer
      2017年度生命科学系学会合同年次大会(ConBio2017)
    • Related Report
      2017 Annual Research Report
  • [Remarks] Vero細胞の物語

    • URL

      https://www.niid.go.jp/niid/ja/chlamydia-pneumonia-m/818-biochem/5752-vero.html

    • Related Report
      2020 Annual Research Report
  • [Remarks] Vero細胞の物語 ~その樹立からゲノム構造の決定、そして未来へ~

    • URL

      http://www.niid.go.jp/niid/ja/chlamydia-pneumonia-m/818-biochem/5752-vero.html

    • Related Report
      2018 Annual Research Report
  • [Remarks] 動物細胞培養に関する用語など

    • URL

      http://www.niid.go.jp/niid/ja/chlamydia-std-m/1880-biochem/3255-2013-02-25-05-58-54.html

    • Related Report
      2018 Annual Research Report
  • [Remarks] Vero細胞の物語 ~その樹立からゲノム構造の決定、そして未来へ~

    • URL

      http://www.nih.go.jp/niid/ja/chlamydia-pneumonia-m/818-biochem/5752-vero.html

    • Related Report
      2017 Annual Research Report
  • [Remarks] 動物細胞培養に関する用語など

    • URL

      http://www.nih.go.jp/niid/ja/chlamydia-std-m/1880-biochem/3255-2013-02-25-05-58-54.html

    • Related Report
      2017 Annual Research Report

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Published: 2017-04-28   Modified: 2023-01-30  

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