Center for functional analysis of human UDN gene homologs in Drosophila and zebrafish

 描述（由申请人提供）：本提案描述了为未诊断疾病网络（UDN）建立模式生物筛选中心（MOSC）。拟议的MOSC将与UDN的中心和研究中心密切合作，在变体选择中发挥作用。领导层将每年从UDN获得300个候选变体的列表，并使用贝勒医学院（Baylor College of Medicine）从罕见疾病队列中获得的独立基因组数据来比较变体，人类表型，并最终每年选择200个变体进行实验。然后，我们将把果蝇中保守的变体（估计每年130个）分配给果蝇核心。在Drosophila Core中，创新技术将用于在内源基因座中用GFP标记蛋白质并检查表达模式。这些品系将通过果蝇基因破坏项目（GDP）的另一项大型合作努力产生，为MOSC节省成本。我们还将使用这种技术来产生突变，这些突变将在一系列表型测定中进行评估。这将指导实验设计和在果蝇中产生携带人类cDNA的转基因。每年分配给俄勒冈州大学的斑马鱼核心的70个估计的变体将通过分析UDN患者的人类表型本体论注释，并利用斑马鱼核心与斑马鱼信息学网络（ZFIN）的连接，使用鱼和小鼠中现有的表达和表型数据进行优先排序。然后，斑马鱼核心将收集现有的突变或用CRISPR/Cas9产生新的突变，并进行高通量表型分析。拟议中的MOSC还计划每两个月与UDN网站和中心共享数据，并建立一个可供整个UDN访问的网站，显示成果和正在进行的工作。不会提及任何个人信息、患者信息或症状，我们只会与直接参与该项目的研究者分享。我们还概述了一个计划，领导与医生互动的诊断或医学上可行的数据。因此，拟议中的中心使用人类基因组学、果蝇和斑马鱼中最具创新性的技术，为UDN患者提供诊断信息。

 DESCRIPTION (provided by applicant): This proposal describes the establishment of a model organism screening center (MOSC) for the Undiagnosed Diseases Network (UDN). The proposed MOSC will play a role in variant selection in close collaboration with the centers and sites of the UDN. The Leadership will obtain from the UDN a list of 300 candidate variants per year, and use independent genomic data available at Baylor College of Medicine (BCM) from rare disease cohorts to compare variants, human phenotypes and ultimately select 200 variants per year for experimentation. We will then assign variants (estimated 130 per year) conserved in Drosophila to the Drosophila Core at BCM. In the Drosophila Core, innovative technology will be used to tag proteins with GFP in the endogenous locus and examine expression pattern. These lines are to be generated by another large collaborative effort through the Drosophila Gene Disruption Project (GDP), saving cost for the MOSC. We will also use this technology to produce mutations that will be evaluated in a battery of phenotypic assays. This will guide experimental design and the generation of transgenics in Drosophila carrying human cDNAs. The 70 estimated variants per year assigned to the Zebrafish Core at the University of Oregon will be prioritized using existing expression and phenotypic data in fish and mice by analyzing the Human Phenotype Ontology annotations of the UDN patients and take advantage of the Zebrafish Core connection to the Zebrafish Informatics Network (ZFIN). The Zebrafish Core will then gathers existing mutations or generate new mutations with CRISPR/Cas9 and perform high throughput phenotypic analyses. The proposed MOSC also plans to share data with the UDN sites and centers every two months and to establish a website accessible to the entire UDN showing results and work in progress. No personal information, patient information or symptoms will be mentioned and we will only share it with investigators that are directly involved in the project. We also outline a plan for the Leadership to interact with the physicians for diagnostic o medically actionable data. Therefore the proposed center uses the most innovative technology in human genomics, Drosophila and zebrafish to provide diagnostic information for UDN patients.