Development of in situ single-cell time-lapse RNA-seq for understanding cell-cell interactions

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K14516
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 28050:Nano/micro-systems-related
• Research Institution: Kyoto University (2023); Institute of Physical and Chemical Research (2021)
• Principal Investigator: Kaneko Taikopaul 京都大学, 医生物学研究所, 助教 (80873504)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 時系列RNA-seq / ナノ局所電気穿孔 / カラーコード / 細胞間相互作用 / 細胞画像と遺伝子発現の統合解析

Outline of Final Research Achievements

Cell-cell interactions are critical factors that temporally influence patterns of gene expression and thereby alter cellular function and fate. However, existing single-cell RNA sequencing techniques only provide snapshots of gene expression at a single time point, leaving the molecular cascades through which cell-cell interactions determine cellular function and fate poorly understood. To address this limitation, we developed in situ single-cell time-lapse RNA-seq, which allows in situ mRNA extraction from living cells and subsequent next-generation sequencing analysis. As elementary technologies, we have established a non-lethal mRNA extraction method using nano-localized electroporation and a color-coding method that allows joint profiling of cellular images and transcriptomes.

Free Research Field

ナノバイオエンジニアリング

Academic Significance and Societal Importance of the Research Achievements

本研究は、静的な遺伝子発現の情報しか得られない既存の１細胞遺伝子解析技術の限界を打破するものであり、これまで得ることが出来なかった遺伝子発現の実時間変動の計測を可能にする点で学術的意義が大きい。本研究で開発するin situ 1細胞タイムラプスRNA-seq法は、例えばがん細胞が免疫細胞から逃れる能力を獲得する過程など、がん細胞の免疫逃避能の分子機序の理解にも役立つと考えられ、その社会的意義も極めて大きい。