Development of opto-molecular genetics by Tet-Light system and its application to sleep-wake regulation

2019 Fiscal Year Final Research Report

• Project/Area Number: 18K19357
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 45:Biology at organismal to population levels and anthropology, and related fields
• Research Institution: University of Tsukuba
• Principal Investigator: Niwa Yasutaka 筑波大学, 国際統合睡眠医科学研究機構, 助教 (40590071)
• Project Period (FY): 2018-06-29 – 2020-03-31
• Keywords: Tet-Lightシステム / Orexin-iCre

Outline of Final Research Achievements

In this study, we tried to construct a Tet-Light system that can temporally control gene expression of the Tet system by photosensitizing tTR, a repressor of the Tet system developed by us. During the research period, we constructed a Tet-Light system using a cultured cell line and were able to clarify the kinetics. We also succeeded in establishing Orexin-iCre knock-in mice, which show in vivo sleep-wake abnormalities and are useful for rescue experiments using the Tet-Light system. In the future, we will use this mouse to verify the in vivo operation and effectiveness of the Tet-Light system.

Free Research Field

睡眠医科学

Academic Significance and Societal Importance of the Research Achievements

個体の行動の中には、神経活動に依存した速い制御だけでなく、睡眠覚醒行動で見られる睡眠負債のようなゆっくりとした細胞生物学的機能の変化がその制御に大きく関わっていると予想されるものも少なくない。そのため従来の光遺伝学による神経活動操作から一歩階層を深めた、遺伝子発現制御を介した細胞内分子の操作、すなわち分子遺伝学的手法が求められている。本研究で構築したTet-Lightシステムは光制御による分子遺伝学的手法を可能にする。今後、Tet-Lightシステムは神経細胞の細胞生物学的機能と個体の行動との間の因果関係を明らかにしていく有効なツールになると考えられる。