斑马鱼肠发育的遗传学与基因组学研究

项目名称： 斑马鱼肠发育的遗传学与基因组学研究

项目编号： No.31471365

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： 黄红辉

作者单位： 西南大学

项目金额： 85万元

中文摘要： 肠是重要的消化器官，研究肠的发育有助于增强对肠相关疾病及出生缺陷的理解。脊椎动物器官发育的机制是保守的，研究斑马鱼肠发育可为人类肠发育提供借鉴。本项目将结合使用遗传学和基因组学方法，系统地研究斑马鱼肠发育的调控基因及机制。我们将开展经典的ENU突变体正向遗传筛选，无基因歧视地鉴定斑马鱼肠发育调控基因；同时我们将通过转录组测序，高效地发掘肠特异性表达基因，通过基因敲除的方法建立突变体并研究其功能与机制。目前，我们已筛选了148个突变基因组，获得了三类共22个肠发育突变体；完成了成鱼全肠及不同节段、胚胎期肠的转录组测序，成鱼肠不同节段的数据分析鉴定了6个组织特异性表达基因簇。这些工作为项目的实施奠定了坚实的基础。我们预期完成400-500个突变基因组的筛选，深入研究2-3个突变体；通过转录组分析鉴定斑马鱼肠特异性表达基因谱，敲除4-5个基因并研究其机制。我们预期可以发现未曾报道的肠发育调控机制。

中文关键词： 斑马鱼；肠发育；正向遗传筛选；转录组；基因敲除

英文摘要： The intestine is an important digestive organ. It is useful to study the intestine development to understand diseases and birth defects related to the intestine in human. Molecular mechanisms of organogenesis among vertebrates are conserved, and results from zebrafish intestine development can be applied to human. To systematically investigate regulatory genes involved in the zebrafsih intestine development; we propose to use both genetic and genomic methods. We are carrying out a classic ENU forward genetic screen for zebrafish intestine mutants to identify genes responsible for the intestine development without gene bias. At the same time, we also perform RNA-seq to investigate transcriptomes of the intestine in adults and embryos, and try to discover specific genes expressed in the intestine, then to study functions of these genes by gene knock-out to generate loss -of -function mutants. We have finished genetic screen of 148 mutagenized genomes and harvested 22 mutants belonging to 3 groups according to phenotypes. We have also collected RNA-seq data of five segments of the adult intestine and the whole intestine in adults and 5dpf embryos. Bioinformatics analyses have identified 6 clads of gene with specific expression patterns in the five segments of the adult intestine and remaining body. These fundamental works have built up a solid foundation for the whole project. By performing the project, we are hoping to finish a screen of 400-500 mutagenized genomes and study 2-3 mutants, identify specific expression genes in the zebrafish intestine and knock-out 4-5 interesting genes. Our study is expected to identify novel molecular mechanisms of the zebrafish intestine development.

英文关键词： zebrafish;the interestine development;forward genetic screen;transcriptome;gene knock-out