Epigenetic mechanisms governing genome partitioning and gene expression in germ cells

控制生殖细胞基因组划分和基因表达的表观遗传机制

• 批准号: 9903402
• 负责人: VALERIE J REINKE
• 金额: $ 41.82万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9903402
• 关键词: 3-Dimensional; Achievement; Binding; Biogenesis; Biological Assay; Bruck-de Lange syndrome; Caenorhabditis elegans; Cell Nucleus; ChIP-seq; Complex; DNA Sequence; Data; Defect; Disease; Elements; Epigenetic Process; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Genomics; Germ; Germ Cells; Germ Lines; Limb structure; Regulation; Regulator Genes; Roberts-SC phocomelia syndrome; Specificity; Structure; System; Translating; Work; X Chromosome; cell type; chromosome conformation capture; experimental study; genetic information; genome-wide; in vivo; in vivo Model; neuronal cell body; piRNA; transcriptome sequencing

Three-dimensional genome organization, in which the linear DNA sequence is partitioned into functionally distinct domains, is essential for the precise deployment of genetic information in every cell type. An increasing number of devastating disorders and diseases have recently been attributed to disruption of genome organization, yet the mechanistic underpinnings remain poorly understood, especially in vivo. In particular, how the activity of binding factors and sequence elements is translated to epigenetic states and three-dimensional genomic domains to govern gene expression remains mysterious. The germ line of C. elegans provides a relevant and experimentally advantageous in vivo model to dissect the mechanisms governing proper establishment of genomic domains with coordinated regulation. In this system, we investigate diverse large-scale gene regulatory mechanisms controlling piRNA biogenesis, X chromosome regulation, and transformation from the germline to the somatic fate. We have recently developed the ability to reliably purify germ nuclei at quantities sufficient for genomic analyses, including ChIP-seq, RNA-seq, and chromatin capture conformation assays. This achievement removes a major barrier in the field to dissect gene expression mechanisms in the germ line. With this new capacity, we will now characterize fundamental transcriptional and epigenetic mechanisms in wild type germ nuclei with unprecedented specificity. Using these data, we will now define key mechanisms governing the germline-to-soma transformation, the coordinated, germline-specific regulation of genomic domains containing thousands of piRNA genes, and the temporal and global regulation of X chromosome gene expression in the germ line. The successful completion of the proposed experiments will illuminate the poorly understood yet vitally important in vivo mechanisms by which large-scale genome organization influences gene expression, and vice versa.

三维基因组组织，其中线性 DNA 序列被划分为 功能不同的域，对于每个细胞中遗传信息的精确部署至关重要 类型。最近越来越多的破坏性病症和疾病被归因于 基因组组织的破坏，但其机制基础仍然知之甚少， 尤其是在体内。特别是，结合因子和序列元件的活性如何转化为 表观遗传状态和控制基因表达的三维基因组结构域仍然存在 神秘。秀丽隐杆线虫种系提供了相关且具有实验优势的体内 模型来剖析控制基因组结构域正确建立的机制 规定。在这个系统中，我们研究了控制多种大规模基因调控机制 piRNA 生物发生、X 染色体调控以及从种系到体细胞命运的转化。 我们最近开发出了可靠纯化胚核的能力，其数量足以用于基因组分析 分析，包括 ChIP-seq、RNA-seq 和染色质捕获构象测定。这个成就 消除了该领域剖析种系基因表达机制的主要障碍。有了这个 新能力，我们现在将表征野生环境中的基本转录和表观遗传机制 具有前所未有的特异性的胚核类型。使用这些数据，我们现在将定义关键机制 控制种系到体细胞的转化，基因组的协调的、种系特异性的调控 包含数千个 piRNA 基因的结构域，以及 X 染色体的时间和全局调控 种系中的基因表达。所提出的实验的成功完成将阐明 大规模基因组组织的体内机制知之甚少但至关重要 影响基因表达，反之亦然。