A Systematic RNAi-based Map of C. elegans Embryogenesis

描述(由申请人提供)：早期线虫胚胎是研究全球遗传结构发育的有力模型。在当前的资助期间，我们已经(1)使用大规模RNAi确定了早期胚胎发育过程中所有可见过程所需的大多数基因，然后分析了延时记录；(2)通过将高分辨率表型与其他功能基因组数据整合起来，建立了早期胚胎发育所需的分子复合体的全局视图；以及(3)通过研究几种新蛋白质的功能来探索图谱的性质。例如，我们发现MEL-28在早期胚胎中发挥着协调核膜和动粒功能的重要作用。在目前的提案中，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，并确定基本复合体是如何相互协调和功能联系的。我们已经开发了一个用于全基因组RNAi的高通量系统，以利用现有的条件等位基因来识别抑制和增强相互作用。利用来自早期胚胎不同过程的代表基因的约30个等位基因，我们的目标是应用这种方法来构建早期胚胎中抑制和增强相互作用的全球支架。我们希望，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背后的复杂表型。 既然人类基因组序列已经被阐明，一个根本的问题是理解它如何在正常和疾病状态下指导复杂的生物学和发育计划。在过去的几年里，我们在破译许多基本发育过程中的单基因需求方面取得了进展，但我们对多基因相互作用如何影响这些程序的看法非常有限。由于大多数人类疾病是由多种相互作用的成分的复杂影响引起的，因此我们必须对这些机制有深入的了解。我们将进行全基因组分析，以揭示在遗传模型线虫的早期发育计划中所需的遗传相互作用。由此得到的图谱将帮助我们了解人类发育和疾病背后的复杂遗传网络。

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.