Regulation of chromatin folding in space and time

染色质折叠在空间和时间上的调节

• 批准号: 10622801
• 负责人: Eric F. Joyce
• 金额: $ 48.75万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622801
• 关键词: Aneuploidy; Architecture; Biological Assay; Cell Nucleus; Cells; Chromatin; Chromatin Loop; Chromosome Positioning; Chromosomes; Complex; DNA; Development; Disease; Enhancers; Fluorescent in Situ Hybridization; Genes; Genetic Transcription; Genome; Genomics; Goals; In Situ; Inherited; Malignant Neoplasms; Methods; Mitotic Recombination; Molecular; Nuclear; Positioning Attribute; Prevention; Proteins; Public Health; Regulation; Resolution; Technology; Time; Work; candidate identification; gene function; genetic information; new therapeutic target; novel; three dimensional structure; tool; Aneuploidy; Architecture; Biological Assay; Cell Nucleus; Cells; Chromatin; Chromatin Loop; Chromosome Positioning; Chromosomes; Complex; DNA; Development; Disease; Enhancers; Fluorescent in Situ Hybridization; Genes; Genetic Transcription; Genome; Genomics; Goals; In Situ; Inherited; Malignant Neoplasms; Methods; Mitotic Recombination; Molecular; Nuclear; Positioning Attribute; Prevention; Proteins; Public Health; Regulation; Resolution; Technology; Time; Work; candidate identification; gene function; genetic information; new therapeutic target; novel; three dimensional structure; tool

Abstract Eukaryotic genomes encode genetic information in their linear sequence, but appropriate expression of their genes requires chromosomes to fold into complex and spatially distinct three-dimensional structures. Recent advances in genomic-based approaches have uncovered a hierarchy of DNA interactions, from small chromatin loops that connect genes and enhancers to larger chromosomal domains and nuclear compartments. However, despite the remarkable conservation of these organizational features and their impact on gene function, we have a very limited understanding of how chromosomes are spatially partitioned, functionally packaged, and relatively positioned in the nucleus. Technical limitations have also hindered our ability to ask questions regarding cell-to-cell variability and the relationship between chromatin folding, positioning, and function at single cell resolution. Our previous studies involved the development of two technologies that use fluorescent in situ hybridization (FISH) to interrogate chromosome positioning at single-cell resolution. Our goal is to build on this work and use these tools to elucidate how chromosomal segments find each other and then form stable interactions within cells. To this end, we have developed a rapid and precise method for identifying candidates involved in chromosome interactions. We now propose to employ this technology to isolate novel architectural proteins, which will be followed by a battery of genomic and in situ-based assays to characterize the candidates. Collectively, the studies proposed here will uncover novel molecular mechanisms underlying nuclear organization, providing a new avenue to study how chromatin folding and positioning is established and inherited, and how dysfunctional organization contributes to disease.

抽象的 真核基因组用线性序列编码遗传信息，但适当的表达 它们的基因需要染色体折叠成复杂和空间不同的三维结构。 基于基因组的方法的最新进展已发现了小型DNA相互作用的层次结构 将基因和增强子连接到较大染色体结构域和核的染色质环路 车厢。然而，尽管这些组织特征及其具有显着的保护 对基因功能的影响，我们对染色体的空间分区方式有非常有限的了解， 功能包装，相对位于核中。技术限制也阻碍了我们的 能够提出有关细胞间变异性以及染色质折叠之间关系的问题， 定位，并在单细胞分辨率下功能。 我们以前的研究涉及开发两种使用荧光原位的技术 杂交（FISH）以单细胞分辨率询问染色体定位。我们的目标是建立在此基础上 工作并使用这些工具来阐明染色体片段如何相互找到，然后形成稳定 细胞内的相互作用。为此，我们开发了一种快速而精确的方法来识别候选人 参与染色体相互作用。我们现在建议采用这项技术隔离新型建筑 蛋白质，随后将是基因组和原位测定的蛋白质，以表征 候选人。总的来说，这里提出的研究将发现基本的新分子机制 核组织，提供了研究如何建立染色质折叠和定位的新途径 并继承，以及功能失调的组织如何对疾病贡献。