Deciphering mechanisms governing functional partitioning of the C. elegans genome

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 神秘的。线虫提供了一种实验上有利的体内模型来解剖 控制基因组结构域正确建立的机制。手头的数据指向一本小说 序列特异性因子 (SNAP190) 与 RNA 成分相互作用的机制 聚合酶 III 复合物定义了一个大的基因组结构域，表现出组织特异性、协调性 数千个非编码小RNA的调控。在这个提案中，强大的遗传和基因组 线虫工具将用于研究 SNAP190、RNA 聚合酶 III、粘连蛋白和 其他候选因素，与染色质状态（目标 1）、染色体环（目标 2）和顺式相互作用 元素（目标 3）来定义这个独特基因组的边界并调节其中的基因表达 域。所提出的实验的成功完成将阐明目前尚知之甚少的问题 驱动体内大规模基因组组织的至关重要的机制。

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. C. elegans provides an experimentally advantageous in vivo model to dissect the mechanisms governing proper establishment of genomic domains. Data in hand points to a novel mechanism in which a sequence-specific factor (SNAP190) acts with components of the RNA polymerase III complex to define a large genomic domain exhibiting tissue-specific, coordinated regulation of thousands of noncoding small RNAs. In this proposal, the powerful genetic and genomic tools of C. elegans will be employed to investigate how SNAP190, RNA polymerase III, cohesin, and other candidate factors, interact with chromatin states (Aim 1), chromosomal looping (Aim 2), and cis- elements (Aim 3) to define the boundaries of, and regulate gene expression within, this unique genomic domain. The successful completion of the proposed experiments will illuminate the poorly understood yet vitally important mechanisms driving large-scale genome organization in vivo.