2021 Fiscal Year Final Research Report
Elucidation of molecular mechanism of epigenetic memory of FGF21 gene and its functional significance in vivo.
| Project/Area Number |
19K09018
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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 54040:Metabolism and endocrinology-related
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| Research Institution | Dokkyo Medical University |
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Principal Investigator |
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | CRISPR-dCas9-TET1CD系 / 遺伝子特異的DNA脱メチル化 / FGF21 / PPARα / エピジェネティクス / エピゲノム制御 / エピゲノム記憶 |
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| Outline of Final Research Achievements |
Recently, we reported PPARα-dependent DNA demethylation of the Fgf21 promoter in the postnatal mouse liver, where reduced DNA methylation is associated with enhanced gene expression after PPARα activation. However, there is no direct evidence for the effect of site-specific DNA methylation on gene expression. We employed the dCas9-SunTag and single-chain variable fragment (scFv)-TET1 catalytic domain (TET1CD) system to induce targeted DNA methylation of the Fgf21 promoter both in vitro and in vivo. We succeeded in targeted DNA demethylation of the Fgf 21 promoter both in Hepa1-6 cells and PPARα-deficient mice, with increased gene expression response to PPARα synthetic ligand administration and fasting, respectively. This study provides direct evidence that the DNA methylation status of a particular gene may determine the magnitude of the gene expression response to activation cues.
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| Free Research Field |
内分泌代謝学
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| Academic Significance and Societal Importance of the Research Achievements |
遺伝子特異的にDNA脱メチル化を導入するエピゲノム改変の研究は世界的にもまだ少なく、本研究は先駆的である。CRISPR-dCAS9-TET1CD系を用いてFGF21遺伝子特異的なDNA脱メチル化を細胞とマウス肝臓に導入した。本研究によりFGF21遺伝子のDNA脱メチル化によるエピゲノム制御の機能的意義が初めて明らかになった。またCRISPR-dCAS9-TET1CD系とPPARαノックアウトマウスを用いた「遺伝子特異的エピゲノム改変動物」は代謝分野において世界で初めての試みである。今後このエピゲノム改変動物を解析することにより、代謝におけるエピゲノム記憶の解明につながることが期待される。
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