Biophysical, Topological, and Functional Studies of Endogenous Circular DNAs

内源性环状 DNA 的生物物理、拓扑和功能研究

• 批准号: 10799325
• 负责人: STEPHEN D. LEVENE
• 金额: $ 20万
• 依托单位: UNIVERSITY OF TEXAS DALLAS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-05 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10799325
• 关键词: Biogenesis; Biological; Biological Process; Biophysics; Cells; Chromatin; Chromosomes; Circular DNA; DNA; Diagnostic; Drug resistance; Elements; Explosion; Genome; Genomic approach; Heterogeneity; Human; Human Cell Line; Human Engineering; Maintenance; Malignant Neoplasms; Molecular; Nucleic Acids; Nucleosomes; Organism; Outcome; Physical Chemistry; Plants; Play; Property; Research; Role; Structure; Techniques; Therapeutic; Variant; Work; biophysical techniques; cancer therapy; chemotherapy; experience; genetic analysis; innovation; personalized medicine; tool; tumor progression

Biophysical, Topological, and Functional Studies of Endogenous Circular DNAs (PI: S.D. Levene) Project Summary/Abstract The existence of endogenous circular DNA elements, termed extrachromosomal circular DNA (eccDNA), has been established in a wide range of organisms from plants to humans for close to fifty years. Until recently, little was known about the functions, biogenesis, and sequence properties of these molecules. However, a recent explosion in the field has begun to amass evidence that the mechanisms producing these species play important roles in genome dynamics. The connection between eccDNA biogenesis and variations in DNA- locus copy number generate cellular heterogeneity, which has important implications for healthy and dysfunctional cells and major impact on cancer progression and treatment. This application offers an unprecedented opportunity to fuse biophysical and genomic approaches that are crucial to advance eccDNA research and builds on the PIs nearly thirty years of experience in DNA topology and nucleic-acid physical chemistry. We will use an innovative, multi-faceted approach that combines biophysical and DNA-topology- based techniques for DNA/chromatin isolation and characterization with leading-edge tools for genetic analysis. Using standard and engineered human cell lines, we will define the roles of eccDNA topology (e.g., supercoiling), sequence, size, and nucleosomal organization in the biological activities of endogenous circular DNAs. A major outcome of this work will be an advanced quantitative understanding of eccDNA biogenesis, maintenance, organization, and biological function. In addition, we will identify structural properties unique to eccDNA elements, which can be exploited for molecular therapeutics and other applications including diagnostics.

内源性环状DNA的生物物理、拓扑和功能研究 (PI：S.D. Levene） 项目总结/摘要 内源性环状DNA元件，称为染色体外环状DNA（eccDNA）， 在从植物到人类的广泛生物体中建立了近50年。直到最近， 对这些分子的功能、生物起源和序列特性知之甚少。但 最近在该领域的爆炸已经开始积累证据，产生这些物种的机制发挥作用， 在基因组动态中的重要作用。eccDNA生物发生与DNA变异之间的联系 基因座拷贝数产生细胞异质性，这对健康和 功能失调的细胞和对癌症进展和治疗的主要影响。该应用程序提供了一个 融合生物物理和基因组方法的前所未有的机会，这对推进eccDNA至关重要 研究和建立在PI近三十年的DNA拓扑学和核酸物理学经验的基础上 化学.我们将使用一种创新的，多方面的方法，结合生物物理和DNA拓扑结构， 基于技术的DNA/染色质分离和表征与领先的工具，遗传 分析.使用标准和工程化的人细胞系，我们将定义eccDNA拓扑结构的作用（例如， 超螺旋），序列，大小和核小体的组织在内源性循环的生物活动 DNA这项工作的一个主要成果将是对eccDNA生物发生的进一步定量理解， 维持、组织和生物功能。此外，我们还将确定独特的结构特性， eccDNA元件，其可用于分子治疗和其它应用，包括 诊断