RBM4调控RNA选择性剪接的机制及其在肿瘤进展中的作用研究

项目名称： RBM4调控RNA选择性剪接的机制及其在肿瘤进展中的作用研究

项目编号： No.31471235

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 汪洋

作者单位： 大连医科大学

项目金额： 85万元

中文摘要： 人类基因发生RNA选择性剪接产生蛋白质的多样性。剪接异常可导致肿瘤，而剪接因子与肿瘤关系尤为密切。我们研究显示剪接因子RBM4能够抑制剪接，并参与调控肿瘤进展过程。本研究拟从前期工作基础出发，采用本实验室经典的GFP报告基因系统（Nat Struct Mol Biol, 2012 & 2013），人工合成剪接因子技术（Nature Methods, 2009）和高通量的RNA-Seq技术系统性研究RBM4调控剪接的类型；鉴定其功能结构域及与RNP复合体相互作用的蛋白结构域，并通过多种分子生物学手段进行验证，以期揭示RBM4调控选择性剪接的机制。在此基础上，本研究还将探讨RBM4通过调控选择性剪接而影响肿瘤进展的生物学功能和分子机制。本研究的顺利实施将为RBM4的研究开辟一个崭新的领域，进一步完善RNA选择性剪接调控的分子网络，并将为人们更好地研究选择性剪接调控与肿瘤提供更加充分的分子基础。

中文关键词： 基因表达调控；mRNA选择性剪接；剪接因子；RBM4；RNA－Seq

英文摘要： In the human genome, more than 90% of all human genes undergo alternative splicing, which allows them to produce multiple splicing isoforms from a single gene locus, thus generating the protein diversity. However, dysregulation of alternative splicing is closely associated with different human diseases, including human cancer. Specifically, protein splicing factors play important roles in cancer progression. Our previous data showed that RBM4, a splicing factor, can inhibit alternative splicing, and is also involved in tumorigenesis. More importantly, there is no related report yet. Based on our preliminary results, we here propose to use the classic GFP reporter gene system, the engineered splicing factors, and the high throughput RNA-sequencing technology to systematically investigate how RBM4 regulates alternative splicing,to identify the functional domains of RBM4 in splicing regulation, and the protein domains of RBM4 in interacting with RNP complex. In addition, we will apply various experimental approaches to validate these findings, thus to reveal the splicing regulation mechanisms of RBM4. Furemore, we will study how RBM4 affects tumor progression through splicing regulation. Collectively, this study will uncover a novel field of RBM4, help us better understand the RNA splicing regulation networks. Additionally, it will provide more molecular basis to study splicing dysregulation and cancer. Therefore, this study have both therotical and technical novelty.

英文关键词： gene expression regulation;mRNA alternative splicing;splicing factor;RBM4;RNA-Seq