elavl1a在斑马鱼心脏发育中的调控机制研究

项目名称： elavl1a在斑马鱼心脏发育中的调控机制研究

项目编号： No.31501164

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 李西

作者单位： 温州医科大学

项目金额： 20万元

中文摘要： Elavl1是一种RNA结合蛋白，对于胚胎的早期发育至关重要。然而，elavl1在早期心脏发育的研究甚少。我们前期工作表明敲降elavl1a斑马鱼胚胎中心脏发育不正常，同时nkx2.5表达下降。这个表型跟gata5突变体非常相似。生物信息学分析发现gata53’UTR存在三个elavl1a的可能结合位点。因此，我们假设elavl1a对心脏的发育具有至关重要的作用；其可能通过直接结合gata5 3’ UTR，保护mRNA稳定性，从而调控gata5的表达。本课题拟在elavl1a敲除的斑马鱼和细胞系中验证elavl1a可以直接结合gata5非编码区，增加 gata5的表达，保证心脏正常发育。本课题是第一次证明elavl1a在心脏发育中的作用，为先天性心脏病的治疗提供一定的理论基础。

中文关键词： 斑马鱼；胚胎发育；gata5；RNA；结合蛋白

英文摘要： Elavl1, an RNA binding protein, is essential for embryonic development. However, its role in zebrafishcardiac development has not been studied. Our preliminary data has shown cardiac looping defect and decreased nkx2.5 expression in the elavl1a knock-downmorphant, which is similiar to gata5 mutant phenotype. Interestingly, zebrafish gata5 3’UTR region contains 3 potential elavl1a binding sites through bioinformatic analysis. Therefore, we hypothesize that elavl1a is essential for cardiac development; mechanistically, elavl1a directly interacts with gata5 3’UTR region, protectgata5 mRNA stability and enhance gata5 expression. In the proposed research project, we will validate cardiac phenotype and marker genes in CRISPR-Cas9 mediated mutants. We will elucidate the association between RNA binding protein elavl1a and gata5 3’UTR in vitro and in vivo by RNA-IP. Furthermore, we will investigate ELAVL1 regutes mRNA stability in lentivirus-mediated ELAVL1knock-down HEK293 cell lines. Thereby, our aim is to uncover a novel role of elavl1a in regulation of cardiac development by protecting gata5 mRNA stability, which provide a signaling pathway for understanding congenial heart disease.

英文关键词： zebrafish;embryonic development;gata5;RNA binding protein