Establishment of artificial control of DNA methylation for stabilization of transgene expression through generations

2023 Fiscal Year Final Research Report

• Project/Area Number: 19K06460
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42040:Laboratory animal science-related
• Research Institution: Osaka Medical and Pharmaceutical University
• Principal Investigator: Yamamoto Yasuhiro 大阪医科薬科大学, 医学部, 講師 (20613558)
• Project Period (FY): 2019-04-01 – 2024-03-31
• Keywords: Transgenerational / DNA methylation

Outline of Final Research Achievements

High DNA methylation of the transgene promoter is thought to be the cause of silencing of the transgene expression state over generations. We analyzed the variation in DNA methylation patterns during medaka gametogenesis and early development by immunohistological methods using 5-methylcytosine antibody, which is specific for DNA methylation. Furthermore, we analyzed which gene groups undergo DNA methylation variation based on comprehensive analysis of DNA methylation status using next-generation sequencers. The results showed that DNA methylation decreased during the egg formation process, suggesting that gene silencing occurs when genes introduced from the mother are transmitted to the next generation.

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Free Research Field

生殖内分泌

Academic Significance and Societal Importance of the Research Achievements

導入遺伝子の安定した発現維持は基礎研究にも応用研究にも関わる重要な事項であり、世代を越えると導入遺伝子の発現が抑制される“導入遺伝子サイレンシング”は解決すべき大きな問題である。導入遺伝子のサイレンシングはプロモーターの高メチル化に起因すると考えられているが、これら因果関係を直接解析した報告は無いため、サイレンシングを回避する効果的な方法は未だ確立されていない。本研究では、導入遺伝子のDNAメチル化を人為的に制御することで、サイレンシングを回避する方法の確立を目的とした。