Kurita Hisaka, Aiba Toshiki, Saito Toshiyuki, Ohsako Seiichiroh
Laboratory of Environmental Health Science, Center for Disease Biology and Integrative Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655 Japan.
Laboratory of Medical Therapeutics and Molecular Therapeutics, Gifu Pharmaceutical University, Daigaku-Nishi, Gifu, 501-1196 Japan.
Genes Environ. 2018 Jan 10;40:1. doi: 10.1186/s41021-017-0089-5. eCollection 2018.
Environmental factors stimulate alteration of DNA methylation level. Investigation of the genome-wide DNA methylation status is important for environmental health studies. We here designed a genomic DNA amplification and labeling protocol using a methylation-sensitive restriction enzyme I. This method can specifically amplify genomic DNA fragments possessing methyl-CpG at the end. The fragments are a relatively short size and dominantly located on CpG-islands. By using the samples prepared by this method, a dioxin-induced change in the methylation level of the mouse promoter was successfully evaluated using oligonucleotide probes covalently bound onto a glass plate. The method developed in this paper would be useful for other genome-wide analysis platforms for the large scale epigenome-wide association studies (EWAS) including human epidemiological samples.
环境因素刺激DNA甲基化水平的改变。全基因组DNA甲基化状态的研究对于环境健康研究很重要。我们在此设计了一种使用甲基化敏感限制酶I的基因组DNA扩增和标记方案。该方法可以特异性扩增末端具有甲基化CpG的基因组DNA片段。这些片段尺寸相对较短，主要位于CpG岛。通过使用该方法制备的样本，利用共价结合在玻璃板上的寡核苷酸探针成功评估了二恶英诱导的小鼠启动子甲基化水平变化。本文开发的方法将有助于其他全基因组分析平台用于大规模表观基因组全关联研究（EWAS），包括人类流行病学样本。
