2023 Fiscal Year Final Research Report
Deciphering the Mechanism of Foxp3 Expression Control in Vivo Using Transcriptional Dynamics Analysis and CRISPR
| Project/Area Number |
21K07082
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 49070:Immunology-related
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| Research Institution | Kumamoto University |
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Principal Investigator |
Ono Masahiro 熊本大学, ヒトレトロウイルス学共同研究センター, 特任教授 (60447951)
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| Project Period (FY) |
2021-04-01 – 2024-03-31
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| Keywords | Transcription / T cells / Fluorescent Timer / Tocky |
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| Outline of Final Research Achievements |
This study advanced the application of a technique developed by the applicant, called Tocky, which uses a fluorescent Timer protein as a reporter gene to analyze transcriptional dynamics at the single-cell level in vivo, specifically targeting Foxp3 transcription dynamics in T cells during immune responses. This research involved the generation and utilization of mice (Foxp3-Tocky CNS2 KO) with a CRISPR-edited deletion in the CNS2 sequence of Foxp3-Tocky. These mice were employed to study various T cell immune induction models, including steady-state, developmental, and disease contexts such as contact hypersensitivity. The research focused on examining how these cells establish a transcription program in response to immune challenges. Special emphasis was placed on understanding the temporal dynamics of Foxp3 transcription and the regulatory effects of CNS2 sequence deletion, particularly as cells transition into an activated state under antigen stimulation or inflammatory conditions.
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| Free Research Field |
Molecular and Systems Immunology
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| Academic Significance and Societal Importance of the Research Achievements |
１）この研究は、免疫細胞内の転写因子の調節の分子的複雑さを強調し、Foxp3の転写動態に関する重要な洞察を提供します。CNS2調節配列とその遺伝子発現制御の役割に焦点を当てることで、遺伝子および細胞生物学の既存概念を再定義する可能性のある遺伝子調節の基本的な側面を探求します。
２）蛍光タイマータンパクをレポーター遺伝子として使用したTocky 技術、とくにFoxp3-Tockyツールは申請者（小野昌弘）が開発した独自の革新的技術です。この技術を発展させ、生体内の一細胞レベルでの遺伝子発現変化をリアルタイムで追跡するこの方法は、細胞プロセスの動的研究に新たな次元を提供します。
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