Regulation of chromatin folding in space and time

染色质折叠在空间和时间上的调节

基本信息

批准号：
10173179
负责人：
Eric F. Joyce
金额：
$ 25万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2023-06-30
项目状态：
已结题

项目摘要

Original Project 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. I can envision three immediate stages for our work. The first is developing a rapid and precise method for identifying candidates involved in chromosome interactions. The second is establishing a battery of in situ-based assays that can be used to characterize the candidates, and the third is translating our findings from model organisms to humans. 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.

原始项目摘要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。我可以设想我们的工作三个阶段。首先是发展迅速的确定参与染色体相互作用的候选者的精确方法。第二个是建立可用于表征候选人的基于原位的测定的电池，第三个正在翻译我们从模型生物到人类的发现。总的来说，这里提出的研究将发现新的分子核组织的机制，提供了一种新的途径来研究染色质折叠和定位是建立和遗传的，以及功能失调的组织如何对疾病贡献。