多功能基因sox9对发育过程调节的分子遗传机制

项目名称： 多功能基因sox9对发育过程调节的分子遗传机制

项目编号： No.31271356

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： 阎一林

作者单位： 苏州大学

项目金额： 80万元

中文摘要： 斑马鱼有两个sox9 基因,sox9a 和sox9b。我们率先建立了斑马鱼sox9a和 sox9b 突变体,阐明了它们在斑马鱼早期骨骼发育过程起到了关键作用。 通过对sox9a/sox9b单突变体及双突变体和野生斑马鱼的芯片研究分析,我们率先发现了sox9b在视网膜发育调节中的新功能。我们还率先阐明了小RNA(mirn140)通过调控pdgf信号通道调节腭软骨的发育。本项研究将利用高通量深度测序对sox9a、sox9b单突变体及双突变体和野生斑马鱼进行转录组和小RNA分析，分别获得受这两个基因单独或共同调控的靶基因和调节sox9a，sox9b以及它们靶基因的小RNA。利用原位杂交和荧光定量PCR等技术验证sox9a或sox9b靶基因在软骨、视网膜、神经嵴、心脏、听囊以及性腺中特异表达情况。这些研究将全面阐明sox9基因功能的分子遗传机制。

中文关键词： sox9；kcnj2；斑马鱼；心脏；安第斯综合征

英文摘要： Zebrafish has two co-orthologs of sox9 genes. We have first generated the zebrafish sox9a and sox9b mutants and revealed clearly the distinct roles of the duplicate genes in skeletal development of zebrafish larva. By microarray analysis on zebrafish sox9 mutants, we have discovered the new function of sox9b in retina development of zeberafish. Also we had successfully identified the first miRNA (mirn140) involved in skeletal facial morphogenesis through modulating Pdgf signaling during midfacial development of zebrafish. Here, we propose to perform transcriptome analysis, including analysis of both messenger RNA (mRNA) and micro-RNA (miRNA) on our zebrafish sox9a and sox9b single and double mutants to compare RNA expression profiles of wild type. This study will allow us to identify genes downstream of sox9 and miRNAs essential for gene regulation related to sox9 function. Also we will find out what genes or microRNAs regulate sox9 gene expression. Then we will further evaluate those potential candidate genes and miRNA expression by whole mount in situ hybridization and qPCR in cartilage, retina, neural crest, heart, ear and gonads of wild type, sox9 single and double mutants. Our investigations will help us to understand the molecular and genetic mechanism of sox9 gene regulation.

英文关键词： sox9；kcnj2；zebrafish；heart；ATS