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Optimization of epigenomic modification with regulatory RNAs considering the presence of endogenous non-coding RNAs

考虑内源非编码 RNA 的存在，利用调节 RNA 优化表观基因组修饰

基本信息

批准号：
22710181
负责人：
OHGANE Jun
金额：
$ 2.5万
依托单位：
Meiji University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Young Scientists (B)
财政年份：
2010
资助国家：
日本
起止时间：
2010 至 2011
项目状态：
已结题

项目摘要

Recent accumulating evidences have indicated that various non-coding RNAs(ncRNAs) are involved in region-specific epigenetic regulations. Since many endogenous ncRNAs show tissue/cell-type-dependent expression patterns, it is important to reveal the relationships between tissue/cell-type-dependent expression and epigenetic regulations. Our data indicated that many genes had ncRNAs within promoter regions of tissu/cell-type-specific genes whose DNA methylation status changed by deficiency of ncRNA-related enzymes. The Sall4 gene, which is crucial for stemness of embryonic stem(ES) cells, had an endogenous antisense ncRNA(ASncRNA) overlapping with the promoter region of the protein-coding Sall4 gene. The Sall4 ASncRNA regulated DNA methylation status of the Sall4 promoter independent of double-strand RNA synthesis by Dicer. An ectopic overexpression of the ASncRNA in fibroblast cells resulted DNA demethylation of the Sall4 promoter region. The Sall4 ASncRNA was proven indispensable for maintenance of unmethylated and activated status in ES cells by shRNA knockdown of the endogenous ASncRNA. Our data also indicated that oocyte-specific histone and environmental chemicals were involved in epigenetic regulations. In conclusion, we could identify many ASncRNAs that regulate DNA methylation status in a target-specific manner, and these ASncRNAs are thought as useful for epigenome modification to establish epigenome-related disease model animals and cells.

近年来越来越多的证据表明，多种非编码RNA（ncRNA）参与了区域特异性的表观遗传调控。由于许多内源性ncRNA显示组织/细胞类型依赖性表达模式，因此揭示组织/细胞类型依赖性表达与表观遗传调控之间的关系是重要的。我们的数据表明，许多基因在组织/细胞类型特异性基因的启动子区域内具有ncRNA，这些基因的DNA甲基化状态因ncRNA相关酶的缺乏而改变。Sall 4基因是胚胎干细胞（ES）的关键基因，其内源性反义ncRNA（ASncRNA）与编码蛋白的Sall 4基因的启动子区重叠。Sall 4 ASncRNA调节Sall 4启动子的DNA甲基化状态，而不依赖于Dicer的双链RNA合成。ASncRNA在成纤维细胞中的异位过表达导致Sall 4启动子区域的DNA去甲基化。通过shRNA敲低内源性ASncRNA，证实了Sall 4 ASncRNA对于维持ES细胞中的未甲基化和活化状态是必不可少的。我们的数据还表明，卵母细胞特异性组蛋白和环境化学物质参与表观遗传调控。总之，我们可以鉴定出许多以靶特异性方式调节DNA甲基化状态的ASncRNA，并且这些ASncRNA被认为可用于表观基因组修饰以建立表观基因组相关疾病模型动物和细胞。