A Systematic RNAi-based Map of C. elegans Embryogenesis

6.研究概要/摘要 早期的C。线虫胚胎是一个强大的模型来研究全球遗传结构的基础 发展在目前的资助期间，我们已经（1）确定了所有基因所需的大部分基因， 在早期胚胎发育过程中使用大规模RNAi， 延时记录;（2）建立了早期所需的分子复合物的全局视图 通过整合高分辨率表型与其他功能基因组数据的胚胎发生;和（3） 通过研究几种新蛋白质的功能，探索了该图谱的性质。比如我们 发现MEL-28在协调核膜和动粒功能中起着重要作用 在早期胚胎中。在目前的建议中，我们的目标是扩展这些研究，以确定蛋白质的作用， 超越基本的，并确定基本复合体是如何协调和功能 彼此相连。我们开发了一个高通量的全基因组RNAi系统， 抑制子和增强子的相互作用使用现有的条件等位基因。使用约30个等位基因， 代表性的基因从不同的过程在早期胚胎，我们的目标是应用这种方法来建立 早期胚胎中抑制子和增强子相互作用的整体支架。我们预计 对这些遗传相互作用的鉴定和分析将极大地扩展我们对基因的认识 以及它们如何相互作用以协调生物过程。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。 十七岁叙事 既然人类基因组序列已经被阐明，一个基本的问题是 了解它如何指导正常和发育过程中复杂的生物和发育程序， 疾病状态。在过去的几年里，我们在破译单个基因方面取得了进展， 在许多基本的发展过程中的要求，但我们有一个非常有限的看法如何 多基因相互作用影响这些程序。由于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 机制等我们将进行全基因组分析，以揭示所需的遗传相互作用， 在遗传模型C的早期发育程序中。优雅生成的地图 将帮助我们理解复杂的遗传网络，这些网络是发育和疾病的基础， 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