MAPPING OF CHROMATIN BORDER ELEMENTS IN THE MAMMALIAN GENOME

哺乳动物基因组中染色质边界元素的图谱

• 批准号: 6395894
• 负责人: Michael A. Goldman
• 金额: $ 12.7万
• 依托单位: SAN FRANCISCO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6395894
• 关键词: 3T3 cells; CHO cells; DNA topoisomerases; Mammalia; acetylation; animal genetic material tag; binding sites; chromatin; computer assisted sequence analysis; computer program /software; computer system design /evaluation; deoxyribonuclease I; gene expression; genetic mapping; genetic regulatory element; genetic transcription; genetically modified animals; genome; human genetic material tag; laboratory mouse; northern blottings; nucleic acid probes; polymerase chain reaction; southern blotting

Mammalian chromosomes are organized as a series of supercoilable looped domains about 100 kilobases in length, each tethered at its end to a nuclear matrix. These loops or chromatin domains are the functional units of transcriptional competence in that each independent domain can be assembled either as active chromatin or as inactive chromatin. Our long- term objective is to understand the function of chromatin domains and the elements that comprise them so that we may acquire a complete understanding of the regulation of gene expression and how it may be manipulated for basic research, medical (gene and cancer therapy) and commercial (drug production and delivery) applications. Most studies have focused on regulatory elements in the immediate vicinity of genes, including promoter and enhancer elements, proteins binding to these, and DNA modification around the promoter. It is now abundantly clear that crucial control elements often lie as much as several hundred kilobases from a gene within a structurally-defined unit which we have referred to as the chromatin domain. Few such functional domains have been characterized in any detail, and when they have the approach has generally has generally been a search for one or a few types of regulatory elements. A thorough understanding requires that we map a number of domains in detail, using a battery of methods to identify all of the potentially important regulatory elements in those domains. Although a matrix attachment region, a replication origin and an insulator, capable of dampening the interaction between a promoter in one domain and an enhancer in another, could co-occur in some instances, these elements may still be functionally distinct and not necessarily overlapping. It is only by mapping out all such elements in a number of cases that we expect to make these important distinctions. The detailed mapping of domains will allow us to identify the crucial regulatory elements in those domains, in preparation for future studies in which we plan to test these elements in experimental systems, or to delete or alter them. The initial mapping we propose here will not only allow us to understand and manipulate these particular domains better, but will hopefully reveal general features that will accelerate our ability to map other domains as well.

哺乳动物染色体被组织为一系列超卷曲环状结构 长度约为 100 KB 的结构域，每个结构域的末端都与 核基质。这些环或染色质结构域是功能单元 转录能力的每个独立域可以是 组装为活性染色质或非活性染色质。我们的长期 术语目标是了解染色质结构域的功能和 组成它们的元素，以便我们可以获得完整的 了解基因表达的调控及其可能的方式 用于基础研究、医学（基因和癌症治疗）和 商业（药品生产和运输）应用。 大多数研究都集中在附近的监管要素上 基因，包括启动子和增强子元件，蛋白质结合 这些，以及启动子周围的 DNA 修饰。现在已经很丰富了 显然，关键的控制元件通常有数百个 我们拥有的结构定义单元内的基因的千碱基 称为染色质结构域。很少有这样的功能域 任何细节的特征，当他们有方法时，通常已经 通常是寻找一种或几种类型的监管要素。 彻底的理解需要我们映射许多领域 细节，使用一系列方法来识别所有潜在的 这些领域的重要监管要素。虽然是矩阵 附着区、复制起点和绝缘体，能够 抑制一个结构域中的启动子与增强子之间的相互作用 在另一种情况下，在某些情况下可能同时出现，这些要素可能仍然是 功能不同且不一定重叠。它仅由 在我们期望的许多情况下绘制出所有这些元素 这些重要的区别。域的详细映射将允许 我们确定这些领域的关键监管要素 为未来的研究做准备，我们计划在其中测试这些元素 实验系统，或删除或更改它们。最初的映射我们 在这里提出不仅能让我们理解和操纵这些 特定领域更好，但有望揭示一般特征 也将加快我们绘制其他领域地图的能力。