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: 10364657
• 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/10364657
• 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序列分为 功能上不同的域，对于每个单元格中的遗传信息的精确部署至关重要 类型。最近，越来越多的破坏性疾病和疾病归因于 基因组组织的破坏，但机械基础的理解仍然很少， 特别是体内。特别是，如何将结合因子和序列元素的活性转化为 表观遗传态和三维基因组结构域控制基因表达 神秘。秀丽隐杆线虫的种生细菌在体内提供了相关且具有实验性的优势 剖析管理基因组域的适当建立的机制 规定。在该系统中，我们研究了控制的多种大型基因调节机制 PIRNA生物发生，X染色体调节以及从种系变为躯体命运的转化。 我们最近开发了可靠地以足以对基因组的数量纯化生殖核的能力 分析，包括CHIP-SEQ，RNA-SEQ和染色质捕获构象测定法。这项成就 消除了该领域的主要障碍，以剖析生殖线中的基因表达机制。与此 新容量，我们现在将表征野生的基本转录和表观遗传机制 具有前所未有的特异性的类型细菌核。使用这些数据，我们现在将定义关键机制 管理种系对瘤的转化，基因组的协调生殖特定调节 包含数千种piRNA基因的结构域以及X染色体的时间和全局调节 种系中的基因表达。拟议的实验的成功完成将阐明 大规模基因组组织的体内机制中知名度较低但至关重要 影响基因表达，反之亦然。