Development of new breeding technology for marine fish using full-life cycle aquaculture system and genome editing

• Project/Area Number: 15H02459
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Aquatic bioproduction science
• Research Institution: Kyushu University
• Principal Investigator: MATSUYAMA Michiya 九州大学, 農学研究院, 教授 (00183955)
• Co-Investigator(Kenkyū-buntansha): 北野 載 九州大学, 農学研究院, 助教 (30635008) ; 坂口 圭史 九州大学, 農学研究院, 准教授 (50396280) ; 長野 直樹 宮崎大学, 農学部, 教授 (50437943)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥42,250,000 (Direct Cost: ¥32,500,000、Indirect Cost: ¥9,750,000); Fiscal Year 2018: ¥9,100,000 (Direct Cost: ¥7,000,000、Indirect Cost: ¥2,100,000); Fiscal Year 2017: ¥9,490,000 (Direct Cost: ¥7,300,000、Indirect Cost: ¥2,190,000); Fiscal Year 2016: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000); Fiscal Year 2015: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000)
• Keywords: ゲノム編集 / 育種 / マサバ / 完全養殖 / 攻撃性 / 生殖 / TALEN / カタクチイワシ / CRISPR/Cas9 / CRISPR/Cあｓ9 / 受精卵 / 水産学

Outline of Final Research Achievements

Although chub mackerel is a promising target species for aquaculture, its survival rate is remarkably low due to co-eating during the juvenile season, like bluefin tuna. In this study, we aimed to create a strain of chub mackerel that destroyed AVTR-V1a2, an aggression-related gene, using genome editing technology. As a result, we succeeded in producing 21 individuals in which both alleles of AVTR-V1a2 were destroyed by TALEN (homo KO). In the future, we plan to use these individuals to produce large quantities of fry and evaluate their characters.

Academic Significance and Societal Importance of the Research Achievements

本研究は、完全養殖系のマサバを対象として、攻撃性に関わる遺伝子をゲノム編集技術により破壊することで、共喰い行動を抑え生残率の向上した、養殖により適した形質をもつマサバの品種作製を試みたものである。本研究により、海産魚を対象にしたゲノム編集
を利用した育種の基盤技術が整備された。ゲノム編集食品の流通が解禁された現在、今後、成長産業として有望視されている魚類養殖産業において、育種の加速化が進展することが期待される。

Research Products

• [Journal Article] Comprehensive Experimental System for a Promising Model Organism Candidate for Marine Teleosts (2019)
  • Author(s): Sakaguchi Keiichi、Yoneda Michio、Sakai Noriyoshi、Nakashima Kanako、Kitano Hajime、Matsuyama Michio
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 1-11
  • DOI: 10.1038/s41598-019-41468-8
  • Peer Reviewed / Open Access
• [Journal Article] 魚の性格を変える 九州大学による“おとなしいマサバ開発” (2019)
  • Author(s): 松山倫也, 大賀浩史
  • Journal Title: 月刊養殖ビジネス Volume: 56 Pages: 61-64
• [Presentation] Production of AVTR-V1a2 knockout strain in chub mackerel, Scomber japonicus, by genome editing with TALEN. (2019)
  • Author(s): Sakanoue R, Ohga H, Shibata K, Nagano N, Kitano H, Sakaguchi K, Kuhara S, Tashiro K, Kim S, Nagasako T, Uchida S, Sakuma , Yamamoto T, Gen K, Fujiwara A, Kazeto Y, Kobayashi T, Ohta K, Matsuyama M
  • Organizer: Marine Biotechnology Conference 2019 in Shizuoka
  • Int'l Joint Research
• [Presentation] Luteinizing hormone β-subunit gene knockout in Japanese anchovy (Engraulis japonicus) by TALEN-based genome editing (2019)
  • Author(s): Kai S, Sakaguchi K, Ohta K, Matsuyama M
  • Organizer: Marine Biotechnology Conference 2019 in Shizuoka
  • Int'l Joint Research
• [Presentation] 新奇モデル生物としてのカタクチイワシ－その特徴と将来展望－ (2019)
  • Author(s): 3.坂口圭史，米田道夫，酒井則良，甲斐公士，太田耕平，大賀浩史，松山倫也
  • Organizer: 平成31年度日本水産学会春季大会
• [Presentation] ゲノム編集技術を利用した魚類の育種と利用 (2019)
  • Author(s): 松山倫也
  • Organizer: 令和元年度水産育種情報交換会
  • Invited
• [Presentation] F0 FOUNDER PRODUCTION OF AVTR-V1A2 KNOCKOUT STRAIN (2018)
  • Author(s): Ohga H, Shibata K, Kai S, Sakanoue R, Kitano H, Ohta K, Nagano n, Sakaguchi K, Kuhara S, Tashiro K, Kim SW, Sakuma T, Yamamoto T, Gen K, Fujiwara A, Kazeto Y, Kobayashi T, Matsuyama M
  • Organizer: Aquaculture America 2018
  • Int'l Joint Research
• [Presentation] Spawning induction of chub mackerel by orally administered nafarelin([des-Gly10, 3-(2-naphthyl)-D-Ala6]-GnRHa) (2017)
  • Author(s): Amezawa K, Ohga H, Yazawa R, Takeuchi Y, Matsuyama M, Yoshizaki G
  • Organizer: International Symposium “Fisheries Science for the Future Generations”
  • Int'l Joint Research
• [Presentation] Development of genome editing technology in chub mackerel, Scomber japonicus and knockout of aVTR-V1a2 by TALEN (2017)
  • Author(s): Shibata K, Ohga H, Nagano N, Sakaguchi K, Kitano H, Ohta K, Kuhara S, Tashiro K, Kim S, Sakuma T, Yamamoto T, Gen , Fujiwara A, Kazeto Y, Kobayashi T, Matsuyama M
  • Organizer: International Symposium “Fisheries Science for the Future Generations”.
  • Int'l Joint Research
• [Presentation] Gene knockout of AVTR-V1a2 by TALEN in chub mackerel, Scomber japonicas. (2016)
  • Author(s): 2.Shibata K, Ohga H, Nagano N, Sakaguchi K, Kitano H, Kuhara S, Tashiro K, Kim S, Sakuma T, Yamamoto T, Gen K, Fujiwara A, Kazeto Y, Kobayashi T, Matsuyama M.
  • Organizer: 13th International Symposium on Agricultural, Food, Environmental and Life Sciences in Asia
  • Place of Presentation: Daejeon, Korea
  • Year and Date: 2016-11-09
  • Int'l Joint Research
• [Presentation] Possible role of the leptin in controlling male chub mackerel puberty. (2016)
  • Author(s): 3.Hirata D, Ohga H, Matsumori K, Kitano H, Nagano N, Yamaguchi A, Matsuyama M.
  • Organizer: 13th International Symposium on Agricultural, Food, Environmental and Life Sciences in Asia
  • Place of Presentation: Daejeon, Korea
  • Year and Date: 2016-11-09
  • Int'l Joint Research
• [Presentation] Production of myostatin gene-knockout Japanese anchovies (Engraulis japonicus) using TALEN-based genome editing. (2015)
  • Author(s): Sakaguchi K, Nakashima K, Kitano H, Nagano N, Matsuyama M.
  • Organizer: The 43rd Scientific Symposium of UJNR Aquaculture Panel
  • Place of Presentation: Nagasaki
  • Year and Date: 2015-11-10
  • Int'l Joint Research
• [Presentation] ゲノム編集技術を用いた水産における育種の現状 (2015)
  • Author(s): 松山倫也
  • Organizer: 農業・漁業・食品・エネルギー・環境分野における先端研究施設の利用に関する研究会
  • Place of Presentation: 鳥栖市
  • Year and Date: 2015-11-06
  • Invited