A Systematic RNAi-based Map of C. elegans Embryogenesis

描述（申请人提供）：秀丽隐杆线虫的早期胚胎是研究潜在发育的全球遗传结构的强大模型。在目前的资助期内，我们已经(1)通过对延时记录的分析，利用大规模RNAi确定了早期胚胎发育过程中所有可见过程所需的大部分基因；(2)通过整合高分辨率表型和其他功能基因组数据，构建了早期胚胎发生所需的分子复合物的全局视图；(3)通过研究几种新蛋白的功能来探索该图谱的性质。例如，我们发现MEL-28在早期胚胎中起着协调核膜和着丝点功能的重要作用。在目前的建议中，我们的目标是扩展这些研究，以确定蛋白质在基本蛋白质之外的作用，并确定基本复合物如何相互协调和功能连接。我们开发了一种高通量的全基因组RNAi系统，利用现有的条件等位基因来识别抑制子和增强子的相互作用。利用来自早期胚胎不同过程的代表性基因的约30个等位基因，我们的目标是应用这种方法建立早期胚胎中抑制因子和增强因子相互作用的全球支架。我们期望对这些基因相互作用的识别和分析将极大地扩展我们对早期胚胎中起作用的基因以及它们如何相互作用以协调生物过程的看法。阐明这些机制将有助于我们进一步了解人类疾病的复杂表型。

DESCRIPTION (provided by applicant): The early C. elegans embryo is a powerful model to study the global genetic architecture underlying development. During the current funding period we have (1) identified most of the genes required for all visible processes during early embryonic development using large-scale RNAi followed by analysis of time-lapse recordings; (2) built a global view of the molecular complexes required for early embryogenesis by integrating high-resolution phenotypes with other functional genomic data; and (3) explored the properties of the map by studying the function of several new proteins. For example, we have found that MEL-28 plays an essential role coordinating nuclear envelope and kinetochore functions in the early embryo. In the current proposal we aim to extend these studies to identify the role of proteins beyond the essential ones and to identify how the essential complexes are coordinated and functionally linked with one other. We have developed a high-throughput system for genome-wide RNAi to identify suppressor and enhancer interactions using existing conditional alleles. Using ~30 alleles of representative genes from diverse processes in the early embryo, we aim to apply this approach to build a global scaffold of suppressor and enhancer interactions in the early embryo. We expect that the identification and analysis of these genetic interactions will extend dramatically our view of the genes working in the early embryo and how they interact to coordinate biological processes. Elucidating these mechanisms will help further our understanding of complex phenotypes underlying human disease. A fundamental question, now that the human genome sequence has been elucidated, is to understand how it directs complex biological and development programs in both normal and disease states. Over the last few years we have made progress in deciphering the single gene requirements in many basic developmental processes but we have a very limited view of how multi-gene interactions affect these programs. Since most human diseases arise from complex effects of multiple interacting components it is imperative that we gain insights into these mechanisms. We will conduct genome-wide analyses to uncover genetic interactions required during the early developmental programs in the genetic model C. elegans. The resulting map will help us understand complex genetic networks underlying development and disease in humans.