A Systematic RNAi-based Map of C. elegans Embryogenesis

基于 RNAi 的系统性线虫胚胎发生图谱

基本信息

批准号：
8049422
负责人：
KRISTIN C GUNSALUS
金额：
$ 3.86万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2010-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8049422
关键词：
Address Affect Alleles Architecture Archives Area Award Biological Biological Assay Biological Process Biology Budgets Buffers C. elegans genome Caenorhabditis elegans Cell Polarity Cell division Chromosome Mapping Code Complex Data Data Analyses Data Collection Databases Development Developmental Process Disease Educational process of instructing Embryo Embryonic Development Enhancers Ensure Event Evolution Fertilization Funding Genes Genetic Genetic Models Genetic screening method Genome Goals Human Human Genome Image Kinetochores Laboratories Lead Link Maps Meiosis Mitosis Modeling Molecular Molecular Genetics Nuclear Envelope Organism Outcome Pathway interactions Pattern Phenotype Play Process Program Development Property Proteins RNA Interference Resolution Role Staging System Testing Time Work Yeasts base combinatorial data integration experience functional genomics gene interaction genetic analysis genome sequencing genome-wide human disease in vivo insight programs protein protein interaction scaffold tool

项目摘要

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.

描述（由申请人提供）：早期线虫胚胎是研究发育背后的整体遗传结构的强大模型。在当前资助期间，我们已经 (1) 使用大规模 RNAi 识别了早期胚胎发育过程中所有可见过程所需的大部分基因，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相结合，建立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质的功能来探索图谱的特性。例如，我们发现 MEL-28 在协调早期胚胎中的核膜和着丝粒功能中发挥着重要作用。在当前的提案中，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，并确定基本复合物如何相互协调和功能性联系。我们开发了一种用于全基因组 RNAi 的高通量系统，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。使用来自早期胚胎不同过程的约 30 个代表性基因的等位基因，我们的目标是应用这种方法来构建早期胚胎中抑制子和增强子相互作用的全局支架。我们期望对这些遗传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被阐明，一个基本问题是了解它如何在正常和疾病状态下指导复杂的生物和发育程序。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都是由多种相互作用成分的复杂影响引起的，因此我们必须深入了解这些机制。我们将进行全基因组分析，以揭示线虫遗传模型早期发育过程中所需的遗传相互作用。由此产生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。