DISSECTING THE GENE REGULATORY NETWORK FOR EPIDERMIS

剖析表皮基因调控网络

• 批准号: 7960153
• 负责人: STEVEN Q IRVINE
• 金额: $ 1.68万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-04 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960153
• 关键词: Animal Model; Animals; Behavioral Research; Binding Sites; Cells; Ciona intestinalis; Computer Retrieval of Information on Scientific Projects Database; Development; Disease; Dissection; Dominant-Negative Mutation; Embryo; Environment; Epidermis; Funding; Gene Expression; Grant; Homeobox; Human; Institution; Laboratories; Measures; Methods; Morphogenesis; Phenotype; Reagent; Regulation; Regulator Genes; Regulatory Element; Research; Research Personnel; Resources; Site-Directed Mutagenesis; Source; Syndrome; Transcription factor genes; Transgenes; Transgenic Organisms; United States National Institutes of Health; Work; ascidian; embryo stage 2; gene discovery

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. PROJECT DESCRIPTION The epidermis is the principal interface between an animal and its environment. Development of the epidermis is dependent on a gene regulatory network that controls its differentiation and morphogenesis. Disruption of this regulatory network causes a number of human developmental and disease syndromes. This proposal aims at the dissection of the gene regulatory network for epidermal development in an emerging model organism, the ascidian Ciona intestinalis, taking the homeobox transcription factor gene DllB as a starting point. The research plan is to use transgenic reagents developed in our laboratory to alter expression of the pivotal regulator of epidermal cell specification CiDllB, and measure effects on the gene regulatory network for epidermal development. The specific aims are 1) measure changes in putative target gene expression levels in embryos altered by misexpression of CiDllB from ectodermal to endomesodermal cells; 2) complete a CiDllB dominant negative transgene and examine the embryonic phenotype and effects on putative target gene expression levels; 3) predict binding sites in previously cloned CiDllB regulatory elements and analyze upstream regulation by site-directed mutagenesis; 4) develop methods for cleavage arrest and to separate ectodermal from endomesodermal cells by blastomere isolation in order to discover genes differentially expressed in the ectodermal lineage. These aims will allow us to work out and validate methods for gene regulatory network discovery applicable to proposals for larger projects which we are developing.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 项目描述 表皮是动物与其环境之间的主要界面。表皮的发育依赖于控制其分化和形态发生的基因调控网络。这一调控网络的破坏会导致许多人类发育和疾病综合征。本建议旨在解剖的基因调控网络的表皮发育的一个新兴的模式生物，海鞘海鞘，同源框转录因子基因DllB为起点。 研究计划是使用我们实验室开发的转基因试剂来改变表皮细胞特异性CiDllB的关键调节因子的表达，并测量对表皮发育的基因调控网络的影响。具体目的是1）测量通过从外胚层到内中胚层细胞的CiD 11B的错误表达而改变的胚胎中推定的靶基因表达水平的变化; 2）完成CiD 11B显性负转基因并检查胚胎表型和对推定的靶基因表达水平的影响; 3）预测先前克隆的CiD 11B调节元件中的结合位点并通过定点诱变分析上游调节; 4）开发卵裂阻滞的方法，并通过卵裂球分离将外胚层细胞与内中胚层细胞分离，以发现外胚层谱系中差异表达的基因。这些目标将使我们能够制定和验证适用于我们正在开发的大型项目提案的基因调控网络发现方法。