Targeted DNA demethylation of the Fgf21 promoter by CRISPR/dCas9-mediated epigenome editing
| Autor: | Tetsuya Yamada, Yoshihiro Ogawa, Izuho Hatada, Kazutaka Tsujimoto, Toshiya Tanaka, Kenichi Kawahori, Miho Hamaguchi, Xunmei Yuan, Yuya Nagaoka, Sumiyo Morita, Nozomi Hanzawa, Koshi Hashimoto, Hiroshi Nishina |
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| Rok vydání: | 2019 |
| Předmět: |
Male
lcsh:Medicine Article Epigenesis Genetic Epigenome Mice Gene expression Epigenome editing CRISPR Animals Clustered Regularly Interspaced Short Palindromic Repeats PPAR alpha Obesity lcsh:Science Promoter Regions Genetic Gene Gene Editing Mice Knockout Multidisciplinary DNA methylation Chemistry lcsh:R Ligand (biochemistry) Cell biology DNA Demethylation Fibroblast Growth Factors Mice Inbred C57BL DNA demethylation lcsh:Q CRISPR-Cas Systems |
| Zdroj: | Scientific Reports Scientific Reports, Vol 10, Iss 1, Pp 1-14 (2020) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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