Image-directed nanoscale photo-crosslinking for the study of sub-nuclear structures

用于亚核结构研究的图像引导纳米级光交联

• 批准号: 9896606
• 负责人: LIN CHEN
• 金额: $ 24.75万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-06 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9896606
• 关键词: 3-Dimensional; Alkynes; Binding; Cell Nucleolus; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromatin Fiber; Chromosomes; Coupled; Custom; DNA; DNA Binding; Development; Dyes; Fishes; Fluorescence; Genes; Genomic DNA; Genomics; Goals; Head; Homeostasis; Image; Image Analysis; In Situ; Label; Lasers; Lighting; Link; Methods; Molecular; Nuclear; Nuclear Lamina; Nuclear Structure; Permeability; Photoaffinity Labels; Positioning Attribute; Protocols documentation; Psoralens; Reaction; Research; Resolution; Series; Source; Specificity; Stains; Structure; Technology; Testing; Toxic effect; Ultraviolet Rays; base; cell growth regulation; covalent bond; crosslink; design; experimental study; genetic information; genome-wide analysis; human disease; image guided; imaging modality; imaging probe; mammalian genome; microscopic imaging; nanometer; nanoscale; new technology

Title: Image-directed nanoscale photo-crosslinking for the study of subnuclear structures Mammalian genomes encode genetic information in linear sequence, yet proper expression of cell-specific genes depends on higher order nuclear organization, from the folding of chromosomes to the assembly of chromosomal domains and nuclear compartments. Defining the dynamic assembly, structure and interaction of these sub-nuclear components is critical to understanding the basic mechanisms of cellular functions and regulations. Cutting-edge imaging methods are revealing the details of the nucleus at increasingly higher resolutions; genomic methods such as Hi-C enable genome-wide analysis of chromatin folding and interactions at the molecular level. However, few technologies are available to integrate the imaging and genomic analyses together. The proposed studies aim to develop an image-directed nanoscale photo-crosslinking technology (INPX) to instantly capture genomic DNA in a selected nuclear volume to enable high resolution spatial and temporal studies of sub-nuclear structures at the single cell level. INPX will greatly facilitate the structure and function study of nuclear organization and subnuclear compartments. !

亚核结构研究中的图像导向纳米级光交联 哺乳动物基因组以线性序列编码遗传信息，但细胞特异性的正确表达 基因依赖于更高级的核组织，从染色体的折叠到染色体的组装 染色体结构域和核隔室。定义动态装配、结构和交互 这些亚核成分对于理解细胞功能的基本机制和 规章制度。尖端的成像方法正在以越来越高的速度揭示细胞核的细节 分辨率；基因组学方法，如Hi-C，使染色质折叠和相互作用的全基因组分析成为可能 在分子水平上。然而，几乎没有可用的技术来整合成像和基因组分析 在一起。所提出的研究旨在开发一种图像导向的纳米级光交联技术 (INPX)在选定的核体积中立即捕获基因组DNA，以实现高分辨率的空间和 在单细胞水平上对亚核结构的时间研究。INPX将极大地促进结构和 核组织和亚核区的功能研究。 好了！