Development of a method for spatio-temporal analysis of gene expression based on RNA visualization and bioluminescence imaging techniques

2022 Fiscal Year Final Research Report

• Project/Area Number: 19H02745
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 34020:Analytical chemistry-related
• Research Institution: The University of Tokyo
• Principal Investigator: Yoshimura Hideaki 東京大学, 大学院理学系研究科(理学部), 助教 (90464205)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: バイオイメージング / 生物発光 / RNA / 細胞 / 遺伝子発現

Outline of Final Research Achievements

The present research aims to develop an RNA probe using bioluminescence to monitor time-course localization alteration of the endogenous target RNA in living cells. In an experiment of the probe solution, the probe quickly responds to the addition and degradation of the target RNA reversibly. Then, the probe expression plasmid was induced into living mammalian cells, and bioluminescence imaging of the cells was observed. In this observation, the target RNA, beta-actin mRNA, formed granule-like structures in the cells, to which growth factor stimulation was administered. The granule-like structures were then concentrated in the expanding cell edge region in response to the administrated stimulation. These results demonstrated the performance of the present probe in monitoring the RNA localization alteration accompanied by cellular physiological events.

Free Research Field

分析化学

Academic Significance and Societal Importance of the Research Achievements

本研究の結果から、開発したプローブは生細胞内において標的とする内在性RNAの細胞現象に伴う局在変化を可視化追跡可能な性能を有することが示された。本プローブの特徴として、標的とするRNAの配列に応じて、そのRNAを標的とするようにカスタムメイドにデザインできることがある。すなわち、本プローブの原理を用いて様々なRNAを標的として生細胞内で局在変化を経時観察できるようになる。またRNAは遺伝子発現産物であり、各種網羅的解析により同定されたRNAが標的とする生理機能の発現過程において示す動態解析にも応用可能である。