Analysis of the interaction between transcriptional oscillations and the extracellular environment in multicellular structure formation

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06659
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 44020:Developmental biology-related
• Research Institution: Toho University
• Principal Investigator: Muramoto Tetsuya 東邦大学, 理学部, 准教授 (10612575)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: ライブイメージング / 遺伝子発現振動 / 転写動態 / CRISPR / 光操作

Outline of Final Research Achievements

Live imaging techniques are valuable for analysing the synchronous oscillation in gene expression during the formation of multicellular structures. In this study, efficient methods were established for generating cells specifically for imaging purposes, employing SpCas9-NG and SpRY. These methods overcome the limitation that the three bases following the target sequence of CRISPR/Cas9 must contain two G bases, thereby enabling efficient knock-in of fluorescent proteins even in genomic sequences with a high AT base composition. Additionally, a gene screening method utilising CRISPR libraries has been developed to further our understanding of the fundamental principles governing multicellular structure formation.

Free Research Field

発生生物学

Academic Significance and Societal Importance of the Research Achievements

多細胞体の構築過程では、分化を誘導するシグナルに応じて適切なタイミングで遺伝子発現が制御され、正確に素過程が進行する。この過程で観察される周期的な遺伝子発現を解析するためには、その現象を直接可視化するイメージング技術が必要不可欠である。今回確立された技術は、多細胞体構築の基本原理といった発生生物学の重要な課題の理解に貢献することが期待される。