Study of molecular mechanism of pregnancy using genome editing technology in pig

• Project/Area Number: 17K08054
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Animal production science
• Research Institution: Azabu University
• Principal Investigator: Naomi Kashiwazaki 麻布大学, 獣医学部, 教授 (90298232)
• Co-Investigator(Kenkyū-buntansha): 伊藤 潤哉 麻布大学, 獣医学部, 准教授 (30454143)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ブタ / 妊娠 / 着床

Outline of Final Research Achievements

We clarified that LIF-LIFR-Stat3 signal is indispensable for embryo implantation in mouse using gene manipulated mice. We also tried to optimize genome editing protocol for mouse and pig embryos and then succeeded in production of genome edited pig embryos.

Academic Significance and Societal Importance of the Research Achievements

本研究で，ブタでは胚自身が着床因子であるLIFを発現すること，またLIF受容体も発現していることを明らかにした．この知見は，例えばブタにおける胚移植時にLIFを投与することなどにより，ブタの効率的生産に寄与できる可能性を示した．またゲノム編集技術を用いて効率的にブタ胚の遺伝子改変を行うことができる可能性を示したことから，今後多くの遺伝子改変ブタを作製することで，基礎生物学および医学分野の研究の発展に寄与できると考えられる．

Research Products

• [Journal Article] Phospholipase Cζ (PLCζ) versus postacrosomal sheath WW domain‐binding protein (PAWP): Which molecule will survive as a sperm factor? (2020)
  • Author(s): Nakai Michiko、Ito Junya、Suyama Ayumi、Kageyama Atsuko、Tobari Yasuko、Kashiwazaki Naomi
  • Journal Title: Animal Science Journal Volume: 91 Issue: 1
  • DOI: 10.1111/asj.13345
  • Peer Reviewed / Open Access
• [Journal Article] Successful vitrification of pronuclear-stage pig embryos with a novel cryoprotective agent, carboxylated ε-poly-L-lysine (2017)
  • Author(s): Kamoshita Maki、Kato Tsubasa、Fujiwara Katsuyoshi、Namiki Takafumi、Matsumura Kazuaki、Hyon Suong-Hyu、Ito Junya、Kashiwazaki Naomi
  • Journal Title: PLOS ONE Volume: 12 Issue: 4 Pages: e0176711-e0176711
  • DOI: 10.1371/journal.pone.0176711
  • NAID: 120006223812
  • Peer Reviewed / Open Access
• [Journal Article] Production of sperm from porcine fetal testicular tissue after cryopreservation and grafting into nude mice (2017)
  • Author(s): Hiroyuki Kaneko, Kazuhiro Kikuchi, Nguyen Thi Men, Michiko Nakai, Junko Noguchi, Naomi Kashiwazaki, Junya Ito
  • Journal Title: Theriogenology Volume: 91 Pages: 154-162
  • DOI: 10.1016/j.theriogenology.2016.12.036
  • NAID: 130007023577
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Induced mutations of Leukemia Inhibitory Factor (LIF) in porcine blastocysts by microinjection with RNAs of CRISPR/Cas9 system (2018)
  • Author(s): Risako HAYASHI, Maki KAMOSHITA, Junya ITO, Naomi KASHIWAZAKI
  • Organizer: Asian Australasian Animal Production Congress 2018
  • Int'l Joint Research
• [Presentation] CRISPR/Cas9 によるブタ胚ゲノム編集における核酸注入濃度が発生能に及ぼす影響 (2018)
  • Author(s): 鴨下 真紀・林 理紗子・伊藤 潤哉・柏崎 直巳
  • Organizer: 日本畜産学会第124回大会