The preparation of gene knock out tick by TALEN using nuclease for infectious model

• Project/Area Number: 26660229
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science
• Research Institution: Kagoshima University
• Principal Investigator: TANAKA TETSUYA 鹿児島大学, 農水産獣医学域獣医学系, 准教授 (00322842)
• Co-Investigator(Renkei-kenkyūsha): MOCHIZUKI MASAMI 鹿児島大学, 農水産獣医学域獣医学系, 教授 (90157834)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: マダニ / マダニ細胞 / アクチンプロモータ－ / フェリチンプロモーター / CAGプロモーター / PGKプロモーター / ゲノム / プロモーター領域 / アクチン / チュブリン / ディフェンシン / PGK / フェリチン / 昆虫細胞

Outline of Final Research Achievements

Although tick studies were conducted by gene silencing using RNA interferance for analyzing the gene function of tick, the skills of gene knockeout and gene transgenic for ticks have not been developing at the morment.Therefore, we tried to incorporate foreign gene into the tick cell line for establishing gene enginering. Although the effect of gene plasmid transfeciton into the tick cell line showed high luceferase activity using pmiGLO or pCAGGS-Luceferase,the effect of plasmid transfection into the tick cell line showed low luceferase activity using actin or ferritin pomotor.These studies indicated that the foreign gene incorporated into the tick may make a break through the praparation of gene konockout or trangenic tick.

Research Products

• [Journal Article] Course of induced infection by Eimeria krijgsmannni in immunocompetent and immunodeficient mice (2016)
  • Author(s): Yuina Ono, Makoto Matsubayashi, Hiroaki Kawaguchi, Masashi Tsujio, Masanobu Mizuno, Tetsuya Tanaka, Tatsunori Masatani, Toshihiro Matsui, Tomohide Matsuo
  • Journal Title: Acta Parasitologica Volume: 115(1) Issue: 1 Pages: 211-5
  • DOI: 10.1007/s00436-015-4737-z
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Role of the tumor necrosis factor receptor-associated factor-type zinc finger domain containing protein 1 (TRAFD1) from the hard tick Haemaphysalis longicornis in immunity against bacterial infection. (2016)
  • Author(s): Takechi R, Galay RL, Matsuo T, Maeda H, Kusakisako K, Talactac MR, Mochizuki M, Fujisaki K, Tanaka T.
  • Journal Title: Ticks and Tick-borne diseases Volume: 7 Issue: 1 Pages: 36-45
  • DOI: 10.1016/j.ttbdis.2015.08.002
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A novel C-type lectin with triple carbohydrate recognition domains has critical roles for the hard tick Haemaphysalis longicornis against Gram-negative bacteria. (2016)
  • Author(s): Maeda H, Miyata T, Kusakisako K, Galay RL, Talactac MR, Umemiya-Shirafuji R, Mochizuki M, Fujisaki K, Tanaka T.
  • Journal Title: Developmental & Comparative Immunology Volume: 57 Pages: 38-47
  • DOI: 10.1016/j.dci.2015.12.015
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Impaired cellular immune response to injected bacteria after knockdown of ferritin genes in the hard tick Haemaphysalis longicornis. (2016)
  • Author(s): Galay RL, Takechi R, Umemiya-Shirafuji R, Talactac MR, Maeda H, Kusakisako K, Mochizuki M, Fujisaki K, Tanaka T.
  • Journal Title: Parasitology International Volume: 65 Issue: 3 Pages: 251-257
  • DOI: 10.1016/j.parint.2016.01.007
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Virucidal activity of Haemaphysalis longicornis longicin P4 peptide against tick-borne encephalitis virus surrogate Langat virus. (2016)
  • Author(s): Talactac MR, Yoshii K, Maeda H, Kusakisako K, Hernandez EP, Tsuji N, Fujisaki K, Galay RL, Tanaka T, Mochizuki M.
  • Journal Title: Parasit Vectors Volume: 9 Issue: 1 Pages: 59-59
  • DOI: 10.1186/s13071-016-1344-5
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Improvement of the cryopreservation method for the Babesia gibsoni parasite by using commercial freezing media (2016)
  • Author(s): Kusakisako, K., Masatani, T., Yada Y., Talactac, M. R., Hernandez, E. P., Maeda, H., Mochizuki, M. and Tanaka, T.
  • Journal Title: Parasitol. Int. Volume: In press Issue: 5 Pages: 532-535
  • DOI: 10.1016/j.parint.2016.02.012
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Induction of gene silencing in Haemaphysalis longicornis ticks through immersion in double-stranded RNA (2016)
  • Author(s): Galay, R. L., Hernandez, E. P., Talactac, M. R., Maeda, H., Kusakisako, K., Umemiya-Shirafuji, R., Mochizuki, M., Fujisaki, K. and Tanaka, T.
  • Journal Title: Ticks Tick Borne Dis. Volume: In press Issue: 5 Pages: 813-816
  • DOI: 10.1016/j.ttbdis.2016.03.018
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Functional analysis of recombinant 2-Cys peroxiredoxin from the hard tick Haemaphysalis longicornis (2015)
  • Author(s): Kusakisako K, Masatani T, Miyata T, Galay RL, Maeda H, Talactac MR, Tsuji N, Mochizuki M, Fujisaki K, Tanaka T.
  • Journal Title: Insect Molecular Biology Volume: 25 Issue: 1 Pages: 16-23
  • DOI: 10.1111/imb.12193
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Identification of the Babesia-responsive leucine-rich repeat domain-containing protein from the hard tick Haemaphysalis longicornis. (2015)
  • Author(s): Maeda H, Kurisu K, Miyata T, Kusakisako K, Galay RL, Rio TM, Mochizuki M, Fujisaki K, Tanaka T.
  • Journal Title: Parasitol. Res. Volume: 114 Issue: 5 Pages: 1793-1802
  • DOI: 10.1007/s00436-015-4365-7
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The widely distributed hard tick, <i>Haemaphysalis longicornis</i>, can retain canine parvovirus, but not be infected in laboratory condition (2015)
  • Author(s): Mori H., Tanaka, T. and Mochizuki, M.
  • Journal Title: Journal of Veterinary Medical Science Volume: 77 Issue: 4 Pages: 405-411
  • DOI: 10.1292/jvms.14-0199
  • NAID: 130005067022
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] The vector potential of Haemaphysalis longicornis ticks for Babesia microti parasites under experimental condition (2015)
  • Author(s): Kusakisako, K., Maeda, H., Galay, R. L., Matsuo, T., Tsujio, M., Umemiya-Shirafuji, R., Mochizuki, M., Fujisaki, K. and Tanaka, T.
  • Journal Title: J. Protozool. Res. Volume: In press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Evaluation and comparison of the potential of two ferritins as anti-tick vaccines against Haemaphysalis longicornis (2014)
  • Author(s): Galay, R. L., Miyata, T., Umemiya-Shirafuji, R., Maeda, H., Kusakisako, K., Tsuji, N., Mochizuki, M., Fujisaki, K. and Tanaka, T.
  • Journal Title: Parasit. Vectors Volume: 7（1） Pages: 482-482
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Iron metabolism in hard ticks (Acari: Ixodidae): The antidote to their toxic diet (2014)
  • Author(s): Galay, R. L., Umemiya-Shirafuji, R., Mochizuki, M., Fujisaki, K. and Tanaka, T.
  • Journal Title: Parasitol. Int. Volume: 64（2) Issue: 2 Pages: 182-189
  • DOI: 10.1016/j.parint.2014.12.005
  • Peer Reviewed
• [Presentation] 病原微生物感染に関与するマダニ生物活性分子について (2015)
  • Author(s): 田仲哲也
  • Organizer: 日本衛生動物学会・日本ダニ学会共催 公開シンポジウム
  • Place of Presentation: 法政大学(千代田区、東京都)
  • Year and Date: 2015-09-11
  • Invited
• [Presentation] Virucidal activity of Haemaphysalis longicornis longicin P4 peptide against tick-borne encephalitis virus surrogate Langat virus (2015)
  • Author(s): Melbourne Rio Talactac, 好井健太朗, 辻 尚利, 藤崎幸蔵, 田仲哲也, 望月雅美
  • Organizer: 第158回日本獣医学会学術集会
  • Place of Presentation: 北里大学(八戸市、青森)
  • Year and Date: 2015-09-07
• [Presentation] Knockdown of ferritin genes increases the susceptibility of the hard tick Haemaphysalis longicornis to bacterial infection (2015)
  • Author(s): Tetsuya Tanaka, Remil Linggatong Galay, Rie Takechi, Rika Umemiya-Shirafuji, Melbourne Rio Talactac, Hiroki Maeda, Kodai Kusakisako, Masami Mochizuki, Kozo Fujisaki
  • Organizer: World Association for the Advancement of Veterinary Parasitology Congress 2015
  • Place of Presentation: The Arena and Convention Centre(Liverpool, UK)
  • Year and Date: 2015-08-16
  • Int'l Joint Research
• [Presentation] Decreased resistance of Haemaphysalis longicornis ticks to bacterialchallenge after knockdown of ferritin molecules (2015)
  • Author(s): Remil Linggatong Galay, 武智理恵, 白藤(梅宮)梨可, Melbourne Rio Talactac, 前田大輝, 草木迫浩大, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第23回SADI
  • Place of Presentation: 名取市文化会館(名取市、宮城)
  • Year and Date: 2015-06-26
• [Presentation] フタトゲチマダニ由来ロイシンリッチリピート保有タンパク質(HlLRR)の発現動態とバベシア原虫に及ぼす影響 (2015)
  • Author(s): 田仲哲也, 栗巣孔士, 前田大輝, 宮田 健, 草木迫浩大, Remil Linggatong Galay, Melbourne Rio Talactac, 望月雅美, 藤崎幸蔵
  • Organizer: 第23回SADI
  • Place of Presentation: 名取市文化会館(名取市、宮城)
  • Year and Date: 2015-06-26
• [Presentation] フタトゲチマダニ由来2-Cys型ペルオキシレドキシン (HlPrx2) の抗酸化部位の検索 (2015)
  • Author(s): 草木迫浩大, 正谷達謄, 宮田 健, Remil Linggatong Galay, 前田大輝, 辻 尚利, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第84回日本寄生虫学会大会
  • Place of Presentation: 杏林大学(三鷹市、東京都)
  • Year and Date: 2015-03-21 – 2015-03-22
• [Presentation] The role of ferritin in immunity of the ixodid tick Haemaphysalis longicornis injected with Escherichia coli (2015)
  • Author(s): Remil Linggatong Galay, Rie Takechi, Rika Umemiya-Shirafuji,Melbourne Rio Talactac, Hiroki Maeda, Kodai Kusakisako, Masami Mochizuki, Kozo Fujisaki, Tetsuya Tanaka
  • Organizer: 第84回日本寄生虫学会大会
  • Place of Presentation: 杏林大学(三鷹市、東京都)
  • Year and Date: 2015-03-21 – 2015-03-22
• [Presentation] Identification of the antioxidant active sites of Haemaphysalis longicornis 2-Cys type peroxiredoxin (HlPrx2) (2015)
  • Author(s): Kodai Kusakisako, Tatsunori Masatani, Takeshi Miyata, Remil Linggatong Galay, Hiroki Maeda, Naotoshi Tsuji, Masami Mochizuki, Kozo Fujisaki, Tetsuya Tanaka
  • Organizer: Joint Symposium of the 6th Veterinary Research and the 3rd Veterinary Education among Universities of Veterinary Medicine in East Asia
  • Place of Presentation: 台湾大学(台北、台湾)
  • Year and Date: 2015-02-02 – 2015-02-03
• [Presentation] ラクトフェリンによる抗マラリア原虫機能の解析 (2014)
  • Author(s): 小森園 亮, 柿本麻美子, 田仲哲也, 李 長春, 渡部久実, Ibrahim Hisham, 若林裕之,山内恒治, 宮田 健
  • Organizer: 第19回日本フードファクター学会
  • Place of Presentation: 鹿児島大学(鹿児島市、鹿児島県)
  • Year and Date: 2014-11-08 – 2014-11-09
• [Presentation] Identification and characterization of a C-type lectin like protein from the hard tick Haemaphysalis longicornis (2014)
  • Author(s): Hiroki Maeda, Takeshi Miyata, Kodai Kusakisako, Remil Linggatong Galay, Rika Umemiya-Shirafuji, Masami Mochizuki, Kozo Fujisaki, Tetsuya Tanaka
  • Organizer: The 2nd Sapporo Summer Seminar for One Health
  • Place of Presentation: 北海道大学(札幌市、北海道)
  • Year and Date: 2014-09-24 – 2014-09-25
• [Presentation] The critical functions of ferritins in Haemaphysalis longicornis and their potential as target molecules for tick control (2014)
  • Author(s): Galay Remil Linggatong, 白藤(梅宮)梨可, 宮田 健, 草木迫浩大, 前田大輝, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 北海道大学(札幌市、北海道)
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] フタトゲチマダニ由来2-Cys型ペルオキシレドキシンの機能解析 (2014)
  • Author(s): 草木迫浩大, 正谷達謄, Galay Remil Linggatong, 前田大輝, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 北海道大学(札幌市、北海道)
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] フタトゲチマダニ由来TNF Receptor Associated Factor(TRAF)の同定とその役割について (2014)
  • Author(s): 武智理恵, Galay Remil Linggatong, 前田大輝, 草木迫浩大, 松尾智英, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第157回日本獣医学会学術集会
  • Place of Presentation: 北海道大学(札幌市、北海道)
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] フタトゲチマダニ由来Ｃ型レクチン様蛋白質の動態および機能解析 (2014)
  • Author(s): 前田大輝, 宮田健, 草木迫浩大, Remil Linggatong Galay, 白藤(梅宮)梨可, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第22回分子寄生虫学ワークショップ第12回分子寄生虫・マラリア研究フォーラム合同大会
  • Place of Presentation: 帯広畜産大学(帯広市、北海道)
  • Year and Date: 2014-08-31 – 2014-09-03
• [Presentation] フタトゲチマダニ由来 2-Cys 型ペルオキシレドキシンの動態 (2014)
  • Author(s): 草木迫浩大, 正谷達謄, Remil Linggatong Galay, 前田大輝, 望月雅美, 藤崎幸蔵, 田仲哲也
  • Organizer: 第22回分子寄生虫学ワークショップ第12回分子寄生虫・マラリア研究フォーラム合同大会
  • Place of Presentation: 帯広畜産大学(帯広市、北海道)
  • Year and Date: 2014-08-31 – 2014-09-03
• [Presentation] The potential of recombinant ferritin 1 and ferritin 2 as anti-tick vaccine against Haemaphysalis longicornis (2014)
  • Author(s): Remil Linggatong Galay, Takeshi Miyata, Rika Umemiya-Shirafuji, Hiroki Maeda, Kodai Kusakisako, Masami Mochizuki, Kozo Fujisaki, Tetsuya Tanaka
  • Organizer: TTP8: Ticks and Tick-Borne Pathogens
  • Place of Presentation: Cape Sun hotel(Cape Town, South Africa)
  • Year and Date: 2014-08-24 – 2014-08-29
• [Presentation] The iron-storage and antioxidant functions of two types of ferritin are critical to successful blood feeding and reproduction of the ixodid tick Haemaphysalis longicornis (2014)
  • Author(s): Galay Remil Linggatong, Umemiya-Shirafuji Rika, Maeda Hiroki, Kusakisako Kodai, Mochizuki Masami, Fujisaki Kozo, Tanaka Tetsuya
  • Organizer: The XIIIth International Congress of Parasitoplogy
  • Place of Presentation: Camino Real Hotel（Mexico City,Mexico)
  • Year and Date: 2014-08-10 – 2014-08-15
• [Presentation] Ticks survive at a threshold between engorgement and starvation (2014)
  • Author(s): Kozo Fujisaki, Naotoshi Tsuji, Xuenan Xuan, Rika Shirafuji-Umemiya, Damdinsuren Boldbaatar, Tetsuya Tanaka, Takeshi Hatta, Banzragch Battur, Badgar Battsetseg
  • Organizer: 14th International Congress of Acarology
  • Place of Presentation: 京都テルサ(京都市、京都府)
  • Year and Date: 2014-07-14 – 2014-07-18
  • Invited
• [Book] Vaccine Design: Methods and Protocols (2016)
  • Author(s): Galay, R. L., Miyata, T., Umemiya-Shirafuji, R., Mochizuki, M., Fujisaki, K. and Tanaka, T.
  • Total Pages: 848
  • Publisher: Springer New York
• [Book] RNA Interference (2016)
  • Author(s): Galay, R. L., Miyata, T., Umemiya-Shirafuji, R., Mochizuki, M., Fujisaki, K. and Tanaka, T.
  • Total Pages: 460
  • Publisher: InTech-Open Access Publisher
• [Book] 第2章 免疫の概念（獣医学教育モデル・コア・カリキュラム準拠 獣医免疫学） (2015)
  • Author(s): 田仲哲也、池田輝雄、岡本まり子、松本安喜（編集：池田輝雄、小川健司、松本安喜）
  • Total Pages: 17
  • Publisher: （株）緑書房
• [Book] 第6章 節足動物各論I（ダニ）（獣医学教育モデル・コア・カリキュラム準拠 寄生虫病学) (2014)
  • Author(s): 田仲哲也 （編集：堀井洋一郎、野中成晃、加藤大智、平 恵介、松本 淳、横山直明）
  • Total Pages: 5
  • Publisher: （株）緑書房
• [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/top.html
• [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/research.html