转录因子RFX2调控小鼠精子细胞发育基因转录及表观遗传状态的机理

项目名称： 转录因子RFX2调控小鼠精子细胞发育基因转录及表观遗传状态的机理

项目编号： No.31471349

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 韩春生

作者单位： 中国科学院动物研究所

项目金额： 82万元

中文摘要： 哺乳动物精子发生过程中会出现多种形态各异的细胞；减数分裂完成后，随着染色质的高度凝缩和基因转录沉默，精子细胞的形态变化尤为剧烈。我们前期通过生物信息学分析，发现转录因子RFX2可能调控大量生精细胞高度表达和特异表达基因的转录；基因敲除小鼠表现为精子发生阻滞在球形精子阶段。本申请计划深入研究RFX2在整个基因组范围对基因转录的调控，重点研究其如何影响精子细胞的表观遗传状态。我们将1）比较野生型和RFX2敲除小鼠蛋白编码基因和非编码RNA基因的表达变化；2）研究DNA和组蛋白甲基化的变化，特别关注DNA羟甲基的变化；3）进而鉴定RFX2的直接靶基因，特别注重受其调控的转录因子；4）利用精原干细胞转基因和移植进一步研究受RFX2调控的二级转录因子是否能部分挽救敲除小鼠的缺陷。这些工作的完成，能够揭示转录因子对于表观遗传状态建立和维护的作用，还有助于发现哺乳动物精子发生中基因调控网络中的关键节点。

中文关键词： 精子发生；染色质重构；转录因子；鞭毛；表观遗传

英文摘要： Diverse morphologically different cell types appear during mammalian spermatogenesis; after meiosis, the morphological change of spermatids is especially dramatic, accompanied by the high degree condensation of chromatins and the silencing of transcription of genes. It has been known that many so-called master switch transcription factors play important roles in the differentiation of cells. Previously, through bioinformatics analysis, we found that transcription factor RFX2 may regulate the transcription of multiple genes; the spermatogenesis of its gene knockout mice stopped at the step 12 spermatids. In this application, we propose to carry out in-depth study of how RFX2 regulates gene transcription at the genome-wide scale, focusing on understanding how it contributes to the establishment and maintenance of the epigenetic status of the spermatids. We will first compare the transcriptomes of the spermatids of the wild-type and RFX2 gene knockout mice, including protein coding and non-coding RNA genes; then investigate how the methylation of DNA and histones changes after gene knockout, paying more close attention to the 5-hydroxymethylation of cytosines in the genome; finally identify direct target genes of RFX2, particularly other transcription factors, aiming to the construction of a gene regulatory network. The accomplishment of these researches will strengthen our understanding of how transcription factors are involved in the establishment and maintenance of epigenetic modifications as well as will help us to identify key nodes in the gene regulatory network of mammalian spermatogenesis.

英文关键词： spermatogenesis;chromatin remodeling;transcription factor;flagellum;epigenetic