HIGH-RESOLUTION PHENOTYPIC PROFILING BASED ON THE COMPLEX ARCHITECTURE OF THE C

基于 C 复杂结构的高分辨率表型分析

• 批准号: 8171265
• 负责人: Karen F Oegema
• 金额: $ 0.24万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171265
• 关键词: Architecture; Caenorhabditis elegans; Cell physiology; Collection; Complex; Computer Retrieval of Information on Scientific Projects Database; Embryo; Eukaryota; Funding; Genes; Genetic; Gonadal structure; Grant; Image; Institution; Link; Maps; Mass Spectrum Analysis; Organism; Phenotype; Research; Research Personnel; Resolution; Resources; Source; Sterility; Time; Tissues; United States National Institutes of Health; base

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We profile the collection of C. elegans genes whose inhibition leads to sterility using high-resolution imaging of the syncytial gonad, a complex tissue whose architecture depends on a spectrum of interacting cellular processes. Analysis of 94 visible features in the gonad partitioned the 554 sterile genes into 102 phenotypic classes, predicting functions for 106 of the 116 uncharacterized genes in the collection while also revealing new functions for previously studied genes. We developed a theoretical framework to assess the significance of functional links between specific gene pairs. Using this framework, we integrated our analysis with a prior time-lapse screen in early embryos to generate a network of 819 essential C. elegans genes that can be viewed at multiple levels of functional resolution. Analysis of specific gene sets in the network indicates that multi-parametric phenotypic profiling in a complex tissue in a multi-cellular organism yields functional maps with a resolution equivalent to genetic interaction-based profiling in unicellular eukaryotes. Mass spectrometry was used to validate our prediction (based on phenotype) that an unknown gene (K10D2.4) associated with the APC/C.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们使用合胞性腺的高分辨率成像来分析秀丽隐杆线虫基因的集合，这些基因的抑制导致不育，合胞性腺是一种复杂的组织，其结构取决于一系列相互作用的细胞过程。 对性腺中 94 个可见特征的分析将 554 个不育基因分为 102 个表型类别，预测了集合中 116 个未表征基因中的 106 个基因的功能，同时也揭示了先前研究的基因的新功能。 我们开发了一个理论框架来评估特定基因对之间功能联系的重要性。 使用这个框架，我们将我们的分析与早期胚胎中的先前延时筛选相结合，生成了一个由 819 个重要的秀丽隐杆线虫基因组成的网络，可以在多个功能分辨率水平上进行查看。对网络中特定基因集的分析表明，多细胞生物体复杂组织中的多参数表型分析产生的功能图谱的分辨率相当于单细胞真核生物中基于遗传相互作用的分析。 使用质谱分析来验证我们的预测（基于表型），即未知基因 (K10D2.4) 与 APC/C 相关。