Genomic Regulation Dlx Bi-gene Cluster Simple Chordate

基因组调控 Dlx 双基因簇 简单脊索动物

• 批准号: 6898971
• 负责人: STEVEN Q IRVINE
• 金额: $ 21.6万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6898971
• 关键词: Tunicata; developmental genetics; gene deletion mutation; gene expression; genetic enhancer element; genetic promoter element; genetic regulation; genetic regulatory element; in situ hybridization; polymerase chain reaction; reporter genes; transcription factor

DESCRIPTION (provided by applicant): The DII-A/DII-B bi-gene cluster of the model chordate dona intestinalis is one of the most compact and organizationally simple examples of a developmental gene cluster known. While some of the regulatory interactions within gene clusters have been analyzed, the simple DII-A/DII-B cluster in C. intestinalis offers an opportunity to obtain a more complete picture of the coordinated regulatory architecture of a whole developmental gene cluster. The first specific aim of this project is to construct a dual reporter transgene to assay the regulatory output of each of the two clustered gene promoters simultaneously. This transgene will then form the basis for deletion experiments to identify the major genomic regulatory elements acting on either or both promoters. Along with this work will be tests of individual reporter transgenes that can assess the possible regulatory activity of internal portions of the non-coding DMA, such as the intergenic region. A second aim, which will support the first, is the determination of the endogenous transcript expression pattern using whole mount in-situ hybridization. The third specific aim is to examine the specificity and positional importance of selected regulatory elements by replacing or moving them within the overall construct. One set of experiments will replace one of the basal promoters with a copy of the other to test whether each basal promoter responds differently to the genomic environment. Another set of experiments will move regulatory modules within the cluster to test the importance of position on the regulatory output. C. intestinalis was chosen as the model organism because of extensive genomic and developmental resources, and because they are ideally suited to the rapid assessment of gene regulation using transgenic reporters in a small-scale research setting. The long-term objective is to contribute to the database of types and modes of genomic regulation, which will help in understanding the molecular genetic basis of human developmental disorders.

描述（由申请人提供）： DII-A/DII-B双基因簇的模式chordate dona dona della的是一个最紧凑的和组织简单的例子，已知的发育基因簇。 虽然已经分析了基因簇内的一些调控相互作用，但C.基因组学提供了一个机会，以获得一个更完整的画面协调监管架构的整个发育基因簇。本项目的第一个具体目标是构建双报告基因转基因，以同时检测两个簇基因启动子中每一个的调控输出。 然后，该转基因将形成缺失实验的基础，以鉴定作用于任一或两个启动子的主要基因组调控元件。沿着这项工作的将是测试个别报告转基因，可以评估非编码DMA的内部部分，如基因间区域的可能的调节活性。第二个目标，这将支持第一个，是内源性转录本的表达模式，使用整体安装原位杂交的测定。第三个具体目标是通过在整个构建体中替换或移动所选调控元件来检查它们的特异性和位置重要性。 一组实验将用另一个的拷贝替换一个基础启动子， 测试每个基础启动子是否对基因组环境有不同的反应。另一组实验将在集群内移动调节模块，以测试位置对调节输出的重要性。C.由于广泛的基因组和发育资源，并且因为它们非常适合于在小规模研究环境中使用转基因报告子快速评估基因调控，所以选择了拟南芥作为模式生物。长期目标是为基因组调控的类型和模式数据库做出贡献，这将有助于理解人类发育障碍的分子遗传基础。