Molecular dissection of Topologically Associating Domains (TADs)

拓扑关联域 (TAD) 的分子解剖

• 批准号: 331208046
• 负责人: Dr. Darío Jesús Lupiáñez García, Ph.D.
• 金额: --
• 依托单位: Centro Andaluz de Biología del Desarrollo (CABD)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2020-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/331208046?language=en
• 关键词: Molecular; dissection; Topologically; Associating; Domains

3D folding is an inherent property of the vertebrate genome in order to accommodate the roughly 2m of DNA that a single cell contains in such a tiny space as the nucleus. This folding, far from being a random process, appears to be highly predetermined and strongly influences gene expression. At the subchromosomal scale, our genome is partitioned in self-interacting units called Topologically Associating Domains (TADs), ranging from a few hundreds of base pairs to several megabases. TADs are delimited by so-called boundary regions that isolate them from the rest of the genome, thus promoting contacts between the loci contained within them. It has been proposed that TADs represent a fundamental structural unit of the genome, which is thought to guide regulatory elements to their cognate genes. Recently, the disruption of TADs and their boundaries was shown to be associated to human disorders such as congenital malformations or cancer. Despite the biological relevance of TADs, the essential components required to build such a fundamental structure remain elusive. In this proposal, I aim to molecularly dissect TAD domains and their boundary regions, in order to understand how these structures can be formed and influenced by their genomic context. By combining cutting-edge technologies such as Capture-HiC or CRISPR/Cas to generate mice carrying specific mutations, I am to: 1) evaluate the contribution of TAD boundaries in domain organization; 2) determine the minimal region to confer boundary function and 3) evaluate the impact of genomic context on TAD boundary function. The potential results derived from this proposal would advance in our understanding of the general principles of chromatin organization and how their alteration can originate abnormal phenotypes.

3D折叠是脊椎动物基因组的固有特性，以容纳单个细胞在细胞核这样的微小空间中包含的大约2 m的DNA。这种折叠，远非一个随机的过程，似乎是高度预先确定的，并强烈影响基因表达。在亚染色体尺度上，我们的基因组被划分为称为拓扑关联结构域（TADs）的自我相互作用单元，范围从几百个碱基对到几个兆碱基。TADs由所谓的边界区域界定，该边界区域将它们与基因组的其余部分隔离，从而促进其中所含的基因座之间的接触。已经提出TADs代表基因组的基本结构单元，其被认为将调控元件引导至其同源基因。最近，TADs及其边界的破坏被证明与人类疾病如先天性畸形或癌症有关。尽管TADs具有生物相关性，但构建这种基本结构所需的基本成分仍然难以捉摸。在这个提议中，我的目标是从分子水平上剖析DNA结构域及其边界区域，以了解这些结构是如何形成的，以及它们的基因组背景如何影响这些结构。通过结合尖端技术，如Capture-HiC或CRISPR/Cas来产生携带特定突变的小鼠，我将：1）评估结构域组织中边界的贡献; 2）确定赋予边界功能的最小区域; 3）评估基因组背景对边界功能的影响。从这一提议中得到的潜在结果将促进我们对染色质组织的一般原理以及它们的改变如何引起异常表型的理解。