Platform for light-inducible, biochemical labeling of ribosomes in living single cells for the novel single-cell analytics based on seamlessly connected live cell imaging and transcriptomics

2021 Fiscal Year Final Research Report

• Project/Area Number: 18K19313
• 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: Komatsu Naoki 国立研究開発法人理化学研究所, 脳神経科学研究センター, 基礎科学特別研究員 (30737440)
• Project Period (FY): 2018-06-29 – 2022-03-31
• Keywords: ビオチン化 / 光操作

Outline of Final Research Achievements

We tried to develop a technical element that would contribute the development of new methodologies that enable gene expression analysis for any cells identified by fluorescence live cell imaging while circumventing the need for a cell detachment process. In particular, we tried to develop a light-activated biotinyltransferase that can be activated by light illumination and chemically modify intracellular protein by biotinylation only in cells illuminated. We noticed that tested candidate molecules showed high background of biotinylation before light illumination. To lower the background, we designed other candidate molecules based on different working principles and constructed expression plasmids for the designed molecules. Further characterization and optimization of the candidate molecules will be performed in future.

Free Research Field

細胞生物学

Academic Significance and Societal Importance of the Research Achievements

多細胞からなる組織や細胞集団の性質に関する理解を更に進めていくにあたり、細胞内のmRNAやタンパク質を1細胞レベルかつ網羅的に解析する手法の高度化・多岐化が求められている。生細胞を標識するための、光と化学修飾を組み合わせた新たな手法について、本研究の実施によりその開発基盤を構築することができた。開発を今後進めることにより、多細胞集団を1細胞解析するための新たな手法が構築されることが期待される。