Study of molecular mechanism of complex brain by ferret

• Project/Area Number: 16K18382
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Yuji Tsunekawa 国立研究開発法人理化学研究所, 多細胞システム形成研究センター, 研究員 (80733352)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 複雑脳発生 / de novo Cre KI / 複雑脳におけるAscl1の機能解析 / de novoノックイン技術の開発 / 複雑脳の発生

Outline of Final Research Achievements

The purpose of this project is to understand molecular mechanism of development of the complex gyrencephalic brain. Mice, which is most used model animals to study neural development but they dont't have gyruses in their brain. Because of this fact, I used the ferret as the model animal of this study. Ferret is a samll handy animals which have gyrencephalic brain, however, it is still difficult to apply genome editing technology to ferret. To solve this problem and progress this project, I reported novel de novo targeted gene knock-in techniques mediated by in utero electroporation and Adeno-associated virus (AAV) into the developing and developed complex brain. Further study to understand molecular mechanism of development of the complex gyrencephalic brain will be required by these newly developed technology.

Research Products

• [Journal Article] Developing a De Novo Targeted Knock-in Method Based on in Utero Electroporation Into the Mammalian Brain (2016)
  • Author(s): Tsunekawa, Yuji, Raymond Kunikane Terhune, Ikumi Fujita, Atsunori Shitamukai, Taeko Suetsugu, Fumio Matsuzaki
  • Journal Title: Development Volume: 143 Issue: 17 Pages: 3216-3222
  • DOI: 10.1242/dev.136325
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration. (2016)
  • Author(s): Suzuki K, Tsunekawa Y, Hernandez-Benitez R, Wu J, Zhu J, Kim EJ, Hatanaka F, Yamamoto M, Araoka T, Li Z, Kurita M, Hishida T, Li M, Aizawa E, Guo S, Chen S, Goebl A, Soligalla RD, Qu J, Jiang T, Fu X, ---, Belmonte JC.
  • Journal Title: Nature Volume: 540(7631) Issue: 7631 Pages: 144-149
  • DOI: 10.1038/nature20565
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Developing de novo genome editing meyhods (2017)
  • Author(s): 恒川雄二
  • Organizer: Conbio2017
  • Int'l Joint Research / Invited
• [Patent(Industrial Property Rights)] US provisional application 62/363,164 (2016)
  • Inventor(s): Yuji Tsunekawa
  • Industrial Property Rights Holder: Yuji Tsunekawa
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-07-15
  • Overseas