A Systematic RNAi-based Map of C. elegans Embryogenesis

6.研究摘要/摘要 线虫早期胚胎是研究全球遗传结构的有力模型 发展。在目前的资助期内，我们(1)确定了所有人类所需的大多数基因 利用大规模RNAi技术分析早期胚胎发育过程中的可视过程 延时记录；(2)建立了早期所需的分子复合体的全球视图 将高分辨率表型与其他功能基因组数据相结合的胚胎发生；以及(3) 通过研究几种新蛋白质的功能，探索了MAP的性质。例如，我们 已经发现MEL-28在协调核膜和动粒功能方面起着重要作用 在早期胚胎中。在目前的提案中，我们的目标是扩展这些研究，以确定蛋白质的作用 超越基本的，并确定基本的复合体是如何协调和功能的 彼此相连。我们已经开发了一种高通量的全基因组RNAi系统来识别 利用现有的条件等位基因抑制和增强相互作用。使用~30个等位基因 来自早期胚胎不同过程的代表性基因，我们的目标是应用这种方法来构建 早期胚胎中抑制子和增强子相互作用的全球支架。我们预计， 对这些遗传交互作用的鉴定和分析将极大地扩展我们对基因的看法 在早期胚胎中工作，以及它们如何相互作用来协调生物过程。澄清这些问题 机制将有助于我们进一步了解人类疾病背后的复杂表型。 17.叙事 既然人类基因组序列已经被阐明，一个根本的问题是 了解它如何指导复杂的生物和发育计划，在正常和 疾病状态。在过去的几年里，我们在破译单一基因方面取得了进展。 许多基本开发过程中的需求，但我们对如何 多基因的相互作用会影响这些程序。因为大多数人类疾病都是由复杂的 多个相互作用的组件的影响我们必须深入了解这些 机械装置。我们将进行全基因组分析，以发现所需的遗传交互作用 在遗传模型的早期发育过程中，线虫。生成的地图 将帮助我们了解复杂的遗传网络在人类的发育和疾病中的作用 人类。 1

6. Research Summary / Abstract 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. 1 7. Narrative 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. 1