Analysis of protein-RNA interactions in membraneless organelles

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K16319
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 54010:Hematology and medical oncology-related
• Research Institution: The University of Tokyo
• Principal Investigator: Yamamoto Keita 東京大学, 大学院新領域創成科学研究科, 助教 (70899301)
• Project Period (FY): 2022-04-01 – 2024-03-31
• Keywords: CRISPR／Cas１３ / 膜の無い構造体 / BiFC / NanoBiT / Split-TurboID

Outline of Final Research Achievements

Inside the cells, there are many structures composed of proteins and nucleic acids. Because they exist without membranes, they are called membraneless organelles (MOs). MOs are expected to have a lot of functions, most of which have not been elucidated. To establish the technique to analyze the function of MOs, we combined CRISPR/Cas13 systems and BiFC, NanoBiT, and Split-TurboID, and developed novel methods. We named them BiFC-dCas13, NanoBiT-dCas13, and Split-TurboID-dCas13, respectively. They are useful for understanding the interaction between protein and RNA, which is important for MOs.

Free Research Field

医学

Academic Significance and Societal Importance of the Research Achievements

細胞内には膜のない構造体と呼ばれる蛋白質とRNAの複合体が多数あります。その役割については大部分が未解明です。膜のない構造体の機能を明らかにするため、RNAを標的とするCRISPR/CasシステムのCRISPR/Cas13とBiFC、NanoBiT、Split-TurboIDの三つの技術を組み合わせた新しいシステムを開発し、代表的な膜の無い構造体「パラスペックル」の解析を行いました。