Deciphering mechanisms governing functional partitioning of the C. elegans genome

描述（由申请人提供）： 三维基因组组织，其中线性DNA序列被划分为功能不同的域，对于在每种细胞类型中精确部署遗传信息至关重要。最近，越来越多的破坏性疾病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，特别是在体内。特别是，结合因子和序列元件的活性如何被翻译成表观遗传状态和三维基因组结构域来控制基因表达仍然是个谜。C. elegans提供了一种实验上有利的体内模型来剖析控制基因组结构域正确建立的机制。手头的数据指出了一种新的机制，其中序列特异性因子（SNAP 190）与RNA聚合酶III复合物的组分一起作用，以定义一个大的基因组结构域，该结构域表现出组织特异性，协调调节数千个非编码小RNA。在这个建议中，强大的遗传和基因组工具的C。线虫将被用来研究SNAP 190、RNA聚合酶III、粘着蛋白和其他候选因子如何与染色质状态（Aim 1）、染色体成环（Aim 2）和顺式元件（Aim 3）相互作用，以确定这一独特基因组结构域的边界，并调节其内的基因表达。拟议实验的成功完成将阐明鲜为人知但至关重要的机制，推动大规模基因组组织在体内。

DESCRIPTION (provided by applicant): 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.