Developing new Cre-loxp system for all organism

Research Project

Project/Area Number	18K19311
Research Category	Grant-in-Aid for Challenging Research (Exploratory)
Allocation Type	Multi-year Fund
Review Section	Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research Institution	Institute of Physical and Chemical Research
Principal Investigator	Tsunekawa Yuji 国立研究開発法人理化学研究所, 生命機能科学研究センター, 研究員 (80733352)
Co-Investigator(Kenkyū-buntansha)	宮道和成国立研究開発法人理化学研究所, 生命機能科学研究センター, チームリーダー (30612577)
Project Period (FY)	2018-06-29 – 2020-03-31
Project Status	Completed (Fiscal Year 2019)
Budget Amount *help	¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000) Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000) Fiscal Year 2018: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywords	de novo Cre knock-in / Cre de novo ノックイン / LeaklessCre / LsCreの開発
Outline of Final Research Achievements	Aim of this project is to develop a new technology to apply Cre-loxp systems for all organisms. Newly developed　de novo genome editing technology by applicant has been modified for this project. As a result of two years of research, Cre-loxp systems now can be applicable to animal species such as ferret (animals which we can apply in utero electroporation) during embryonic stages. We father tried to apply Cre-loxp systems to adult animals, however, examined AAV vectors for adult animals caused leak of cre, that was not seen in in utero electroporation system. Further studied is required to fix this newly coming up problem.
Academic Significance and Societal Importance of the Research Achievements	2年間の研究の結果、プラスミドを用いたシステムではマウス、フェレットなどの発生期個体においてCreを導入する技術を確立することができた。この成果を持って特許の申請も行なっている。この新しいシステムを用いることにより、今までは１年以上の時間がかかっていたコンディショナルTgマウスなどが受精卵ノックインなどを用いて簡便に作れる可能性を示しており、産業的に価値があると思われる。