介导人类基因组DNA甲基化修饰的非编码RNA的鉴定及其作用机理研究

项目名称： 介导人类基因组DNA甲基化修饰的非编码RNA的鉴定及其作用机理研究

项目编号： No.31271375

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 李思光

作者单位： 同济大学

项目金额： 80万元

中文摘要： DNA甲基化修饰对干细胞的分化起着重要调控作用。但是在干细胞的分化过程中，DNA甲基化转移酶如何识别特定的DNA位点，催化其甲基化修饰？这个问题迄今没有被解析。我们在前期研究中发现，Dnmt3b复合物中存在非编码RNA,这些非编码RNA可能介导了DNA的甲基化修饰。本项目拟建立人ES细胞向神经细胞分化过程中，与DNMT3复合物结合的RNA 深度测序文库，获得与Dnmt3a和Dnmt3b复合物结合的非编码RNA的序列信息，鉴定出能够介导基因组DNA甲基化修饰的非编码RNA，研究参与DNA甲基化修饰的Dnmt3复合物的组成及各成员在DNA甲基化修饰中所起的作用，揭示非编码RNA介导DNA甲基化修饰的分子机制。本项目的实施不仅能够鉴定一批介导DNA甲基化修饰的非编码RNA，阐明细胞分化的表观遗传调控机制，而且能够建立通过非编码RNA调控DNA甲基化进而诱导ES细胞向神经细胞高效分化的新技术。

中文关键词： DNA甲基化；非编码RNA；细胞分化；；

英文摘要： DNA methlation modification have key regulatory roles in stem cell differentiation. In the process of cell differentiation, how the DNA methyltransferases recognize specific sites in genome for site-specidic DNA methylation is still an open question. Our previous work shows that in the Dnmt3b complexe there are non-coding RNAs (ncRNA) which may direct DNA methlation modification. To investigate the function of ncRNA on the DNA methlation modification during the process of human cell diffrentiation from embryonic stem cell toward neural stem cell, we will isolate RNA from RNA and Dnmt3 complexes and construct the RNA libraries. Then, we will obtain ncRNA sequence from Dnmt3a-ncRNA and Dnmt3b-ncRNA complexes by deep sequencing and identify ncRNAs which direct DNA methlation modification. Finally, we will study the role of members in Dnmt3a-ncRNA and Dnmt3b-ncRNA complexes on the DNA methlation modification and reveal valuable insight into the molecular mechanisms of RNA directing DNA methlation modification.The research of this project will not only identify a group of ncRNAs which direct DNA methlation modification, but also provide valuable method for the differentiation of embryonic stem cell toward neural stem cell by regulating ncRNAs which direct site-specific DNA methylation.

英文关键词： DNA methylation；non-coding RNA；cell differentiation；；