Structural Annotation of the human Genome

人类基因组的结构注释

• 批准号: 7921275
• 负责人: Job Dekker
• 金额: $ 51.27万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-11 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7921275
• 关键词: Affect; Biological; Biological Assay; Carbon; Cell Line; Chromatin Loop; Chromosomes; Data; Data Set; Defect; Detection; Development; Disease; Distal; Distant; Elements; Enhancers; Event; Fluorescent in Situ Hybridization; Gene Expression; Gene Expression Regulation; Gene Order; Gene Targeting; Genes; Genome; Genomics; Gold; Group Identifications; Human; Human Genome; Hypersensitivity; Maps; Methodology; Methods; Molecular Conformation; National Human Genome Research Institute; Occupations; Phase; Positioning Attribute; Prevalence; Process; Regulation; Regulatory Element; Relative (related person); Reporter; Research Personnel; Role; Stretching; Testing; Time; Transfection; base; beta Globin; combinatorial; functional genomics; genetic element; histone modification; insight; novel; programs; promoter; technology development; tool

DESCRIPTION (provided by applicant): Regulation of gene expression is crucial for normal development and defects in this process can result in disease. A better understanding of the genetic elements that contribute to regulation of disease genes can reveal causes of disease and may spur development of novel treatments. Large-scale analyses of genomes, such as those initiated by the ENCODE project, have started to reveal the positions of many regulatory elements. However, in most cases it is not known which elements regulate any given gene. Understanding the relationships between regulatory elements and genes is complicated by the fact that the linear order of elements and genes along chromosomes often does not reflect functional relationships between them. For instance, regulatory elements may affect distal genes but not those located immediately next to them. Therefore, functional connections between genes and regulatory elements must be experimentally determined. This proposal is based on the hypothesis that regulatory elements physically associate with their target gene through formation of chromatin loops. We propose to test this hypothesis by mapping of looping interactions between genes and regulatory elements throughout the well-studied 1 % of the genome selected by the ENCODE consortium. We will detect chromatin loops using a unique approach, Chromosome Conformation Capture (3C) methodology. During the last 2 years we have developed a new high-throughput 3C application, called 5C (for 3C-carbon-copy) which employs microarrays or quantitative sequencing for detection of chromatin loops. We will further optimize 5C by analysis of the beta-globin locus (aim 1). We will employ 5C to identify chromatin loops between genes and regulatory elements throughout the well-studied ENCODE regions of the human genome (aim 2). For each gene we will identify distant elements such as enhancers that interact with its promoter. We will validate looping interactions by FISH (aim 3). We will test the function of looping elements using transient transfections and by integrating looping data with other data obtained by the ENCODE consortium such as histone modifications, DNasel hypersensitivity and target gene expression (aim 3). We will make looping data available through the UCSC genome browser. These studies will map the network of connections between genes and regulatory elements and will reveal new insights into the mechanisms that underlie long-range gene regulation.

描述(由申请人提供)：基因表达调控对正常发育至关重要，这一过程中的缺陷可能会导致疾病。更好地了解有助于疾病基因调控的遗传因素可以揭示疾病的原因，并可能刺激新疗法的开发。对基因组的大规模分析，例如由ENCODE项目发起的那些，已经开始揭示许多调控元件的位置。然而，在大多数情况下，并不知道哪些元件调节任何给定的基因。元件和基因沿染色体的线性顺序往往不能反映它们之间的功能关系，这使得理解调控元件和基因之间的关系变得复杂。例如，调控元件可能会影响远端基因，但不会影响紧邻它们的基因。因此，基因和调控元件之间的功能联系必须通过实验来确定。这一建议是基于这样的假设，即调控元件通过形成染色质环与其目标基因物理联系。我们建议通过绘制ENCODE联盟选择的基因组中经过充分研究的1%的基因组中基因和调控元件之间的循环相互作用来验证这一假设。我们将使用一种独特的方法-染色体构象捕获(3C)方法来检测染色质环。在过去的两年里，我们开发了一种新的高通量3C应用程序，称为5C(即3C-Carbon-Copy)，它使用微阵列或定量测序来检测染色质环。我们将通过分析β-珠蛋白基因座进一步优化5C(目标1)。我们将使用5C来识别基因之间的染色质环和整个人类基因组中研究得很好的ENCODE区域的调控元件(目标2)。对于每个基因，我们将确定与其启动子相互作用的远距离元件，如增强子。我们将通过FISH验证循环交互(目标3)。我们将使用瞬时转染法测试环状元件的功能，并将环状数据与ENCODE联盟获得的其他数据相结合，如组蛋白修饰、DNasel超敏反应和靶基因表达(AIM 3)。我们将通过UCSC基因组浏览器提供循环数据。这些研究将绘制基因和调控元件之间的联系网络，并将揭示奠定基因远程调控基础的机制的新见解。