Development of tick genetically modified drive to prevent pathogen transmission

• Project/Area Number: 17K19328
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science, Animal science, and related fields
• Research Institution: Kagoshima University
• Principal Investigator: TANAKA TETSUYA 鹿児島大学, 農水産獣医学域獣医学系, 教授 (00322842)
• Project Period (FY): 2017-06-30 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: マダニ / ノックダウン / ノックアウト / CRISPR/Cas9 / トランスジェニック / フェリチン / グルタチオンＳトランスフェラーゼ / ペルオキシレドキシン / トランスポゾン / 感染防御分子 / 鉄 / 遺伝子発現 / 遺伝子導入

Outline of Final Research Achievements

We attempted to establish a gene knockout technology related to the protection of ticks and aimed to create an infectious disease model using them. To investigate the involvement of defensins in the viral immune response in ticks, we used hemolymph-derived (HE) defensins and examined their effects on virus growth. When the Langat virus was inoculated in the produced HE defensin knockdown / knockout ticks in the body, it had little effect on the survival of the ticks and the virus multiplication in the body. Therefore, it was considered that the antimicrobial peptide is unlikely to be involved in the antiviral immune response to Langat virus in the body of the tick.

Academic Significance and Societal Importance of the Research Achievements

[学術的意義] マダニの飼育には通常の研究室のような比較的小規模で、何百匹という単位での大規模感染実験が可能である。大量の個体に病原体を感染させられるということは、宿主側の遺伝子変異体を大量に用意することで、病原体に対する感染防御遺伝子の明確な解析を行うことが可能である。
[社会的意義] 哺乳類に比べ、個体そのものを扱った感染実験が容易である。従来のマウスやラットなどの哺乳類を扱った実験系では、1回の実験に経済的や物理的弊害が生じてしまうのが現実である。また、動物を用いた感染実験の倫理面も重視する必要がある。

Research Products

• [Journal Article] Characterization of an iron-inducible Haemaphysalis longicornis tick-derived promoter in an Ixodes scapularis-derived tick cell line (ISE6). (2019)
  • Author(s): Hernandez EP, Kusakisako K, Hatta T, Tanaka T.
  • Journal Title: Parasit Vectors. Volume: 12(1) Issue: 1 Pages: 321-321
  • DOI: 10.1186/s13071-019-3574-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Vector competence of Haemaphysalis longicornis ticks for a Japanese isolate of the Thogoto virus (2018)
  • Author(s): Talactac MR, Yoshii K, Hernandez EP, Kusakisako K, Galay RL, Fujisaki K, Mochizuki M, Tanaka T.
  • Journal Title: Sci. Rep. Volume: 8(1) Issue: 1 Pages: 9300-9300
  • DOI: 10.1038/s41598-018-27483-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hemolymph defensin from the hard tick Haemaphysalis longicornis attacks Gram-positive bacteria (2018)
  • Author(s): Yada, Y., Talactac, M. R., Kusakisako, K., Hernandez, E. P., Galay, R. L., Andoh, M., Fujisaki, K., and Tanaka, T.
  • Journal Title: J. Invertebr. Pathol. Volume: 156 Pages: 14-18
  • DOI: 10.1016/j.jip.2018.07.005
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Induction of intracellular ferritin expression in embryo-derived Ixodes scapularis cell line (ISE6) (2018)
  • Author(s): Hernandez, E. P., Kusakisako, K., Talactac, M. R., Galay, R. L., Yoshii, K. and Tanaka, T.
  • Journal Title: Sci. Rep. Volume: 8(1) Issue: 1 Pages: 16566-16566
  • DOI: 10.1038/s41598-018-34860-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A continuing exploration of tick-virus interactions using various experimental viral infections of hard ticks (2018)
  • Author(s): Talactac, M. R., Hernandez, E. P., Fujisaki, K. and Tanaka, T.
  • Journal Title: Front. Physiol. Volume: 9 Pages: 1728-1728
  • DOI: 10.3389/fphys.2018.01728
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transcriptional activities of two newly identified Haemaphysalis longicornis tick-derived promoter regions in the Ixodes scapularis tick cell line (ISE6) (2018)
  • Author(s): Kusakisako, K., Ido, A., Masatani, T., Morokuma, H., Hernandez, E. P., Talactac, M. R., Yoshii, K. and Tanaka, T.
  • Journal Title: Insect Mol. Biol. Volume: In press
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Synchronous Langat virus infection of Haemaphysalis longicornis using anal pore microinjection (2017)
  • Author(s): Talactac, M. R., Yoshii, K., Hernandez, E. P., Kusakisako, K., Galay, R. L., Fujisaki, K., Mochizuki, M. and Tanaka, T.
  • Journal Title: Viruses Volume: 9（7） Issue: 7 Pages: 189-189
  • DOI: 10.3390/v9070189
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Characterization of an iron-inducible Haemaphysalis longicornis tick-derived promoter in an Ixodes scapularis derived tick cell line (ISE6) (2019)
  • Author(s): Emmauel Pacia Hernandez, 草木迫浩大, 八田岳士, 田仲哲也
  • Organizer: 第88回日本寄生虫学会大会
• [Presentation] 哺乳類細胞によるマダニアクチン遺伝子プロモータを用いた外来遺伝子の発現 (2019)
  • Author(s): 田仲哲也, 諸熊遥樹, 草木迫浩大, Emmanuel Pacia Hernandez
  • Organizer: 第27回ダニと疾患のインターフェイスに関するセミナー
• [Presentation] Evaluation of an iron-inducible Haemaphysalis longicornis tick-derived promoter activity in an Ixodes scapularis-derived tick cell line (2019)
  • Author(s): Emmanuel Pacia Hernandez, Kodai Kusakisako, Takeshi Hatta, Tetsuya Tanaka
  • Organizer: The 7th Sapporo Summer Symposium for One Health
  • Int'l Joint Research
• [Presentation] マダニ胚由来細胞における遺伝子発現ベクターの探索 (2018)
  • Author(s): 田仲哲也, 井戸明子, 草木迫浩大, 諸熊遥樹, 正谷 達謄, Hernandez Emmanuel, Talactac Melbourne, 好井健太郎
  • Organizer: 第26回ダニと疾患のインターフェイスに関するセミナー
• [Presentation] フタトゲチマダニ由来抗酸化蛋白質を発現する哺乳類細胞の安定発現株の作製 (2018)
  • Author(s): 諸熊遥樹, 草木迫浩大, Melbourne Rio Talactac, Emmanuel Pacia Hernandez, 田仲哲也
  • Organizer: 第71回日本寄生虫学会南日本支部大会・第68回日本衛生動物学会南日本支部大会合同大会
• [Presentation] Antibacterial activity by hemolymph defensin from the hard tick Haemaphysalis longicornis (2018)
  • Author(s): Tetsuya Tanaka, Yurika Yada, Melbourne Rio Talactac, Kodai Kusakisako, Emmauel Pacia Hernandez, Remil Linggatong Galay, Masako Andoh, Kozo Fujisaki
  • Organizer: International Symposium in Veterinary Science: Strengthening the Regional Veterinary Education and Research for the Future Excellent Veterinary Graduates
  • Int'l Joint Research
• [Presentation] Evaluation of the vector competency of Haemaphysalis longicornis ticks in transmitting the Japan isolate Thogoto virus to mice (2018)
  • Author(s): Talactac Rio, Kentaro Yoshii, Emmauel Hernandez Pacia, Remil Galay Linggatong, Kodai Kusakisako, Kozo Fujisaki, Masami Mochizuki, Tetsuya Tanaka
  • Organizer: International Symposium in Veterinary Science: Strengthening the Regional Veterinary Education and Research for the Future Excellent Veterinary Graduates
  • Int'l Joint Research
• [Presentation] Investigation of the survival dynamics of Japan isolate Thogoto virus in Haemaphysalis longicornis (2017)
  • Author(s): Melbourne Rio Talactac, Kentaro Yoshii, Emmauel Pacia Hernandez, Kodai Kusakisako, Hiroki Maeda, Kozo Fujisaki, Masami Mochizuki, Tetsuya Tanaka
  • Organizer: International Symposium in Veterinary Science: Strengthening The Collaboration between Indonesian and Japanesse Veterinary School
  • Int'l Joint Research
• [Presentation] Gene silencing in ticks through injection and immersion technique (2017)
  • Author(s): Remil Linggatong Galay, Takeshi Miyata, Tetsuya Tanaka
  • Organizer: 9th PSP Scientific Meeting
  • Int'l Joint Research
• [Presentation] The effect of bacteria by hemolymph defensin from the hard tick Haemaphysalis longicornis (2017)
  • Author(s): Tetsuya Tanaka, Yurika Yada, Melbourne Rio Talactac, Kodai Kusakisako, Emmauel Pacia Hernandez, Hiroki Maeda, Remil Linggatong Galay, Masako Andoh, Masami Mochizuki, Kozo Fujisaki
  • Organizer: One Health 9th Tick and Tick-borne Pathogen Conference and 1st Asia Pacific Rikettsia Conference(TTP9-APRC1)
  • Int'l Joint Research
• [Presentation] Experimental transmission of Japan isolate Thogoto virus by Haemaphysalis longicornis ticks (2017)
  • Author(s): Melbourne Rio Talactac, Kentaro Yoshii, Emmauel Pacia Hernandez, Kodai Kusakisako, Hiroki Maeda, Kozo Fujisaki, Masami Mochizuki, Tetsuya Tanaka
  • Organizer: One Health 9th Tick and Tick-borne Pathogen Conference and 1st Asia Pacific Rikettsia Conference(TTP9-APRC1)
  • Int'l Joint Research
• [Presentation] Characterization and antiviral activity of a newly identified defensin-like peptide, HEdefensin, in the hard tick Haemaphysalis longicornis (2017)
  • Author(s): Melbourne Talactac, 屋田友里花, 好井健太朗, Emmauel Hernandez, 草木迫浩大, 藤崎幸蔵, 望月雅美, Kozo Fujisaki, Masami Mochizuki, 田仲哲也
  • Organizer: 第160回日本獣医学会学術集会
• [Presentation] Evaluation of the vector competency of Haemaphysalis longicornis ticks in transmitting the Japan isolate Thogoto virus to mice (2017)
  • Author(s): Melbourne Talactac, 好井健太朗, Emmauel Hernandez, 草木迫浩大, 藤崎幸蔵, 望月雅美, 田仲哲也
  • Organizer: 第65回日本ウイルス学会学術集会
• [Presentation] 哺乳類細胞におけるマダニアクチン遺伝子プロモーターを用いた外来遺伝子の発現 (2017)
  • Author(s): 諸熊遥樹, 草木迫浩大, Emmanuel Pacia Hernandez, Melbourne Rio Talactac, 田仲哲也
  • Organizer: 第70回日本寄生虫学会南日本支部大会・第67回日本衛生動物学会南日本支部大会合同大会
• [Presentation] マダニ胚由来細胞における遺伝子発現ベクターの探索並びに構築 (2017)
  • Author(s): 田仲哲也, 井戸明子, 草木迫浩大, 諸熊遥樹, 正谷 達謄, Hernandez Emmanuel, Talactac Melbourne, 好井健太郎
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Remarks] 鹿児島大学 共同獣医学部獣医学科 病態予防獣医学講座 感染症学分野
  • URL: http://www.vet.kagoshima-u.ac.jp/kadai/V-Infection/sinkoukansen/index.html