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)建立一个模式生物筛查中心(MOSC)。拟议的MOSC将与统一数字网络的中心和地点密切合作，在变种选择方面发挥作用。领导层将从UDN获得每年300个候选变种的列表，并使用贝勒医学院(BCM)从罕见疾病队列中获得的独立基因组数据来比较变种和人类表型，最终选择每年200个变种进行实验。然后，我们将把果蝇体内保存的变异(估计每年130个)分配给BCM的果蝇核心。在果蝇核心中，将使用创新技术在内源基因座上标记绿色荧光蛋白并研究表达模式。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。我们还将使用这项技术产生突变，这些突变将在一系列表型分析中进行评估。这将指导携带人类cDNA的果蝇的实验设计和转基因基因的产生。俄勒冈大学每年分配给斑马鱼核心的70个估计变体将利用鱼类和小鼠的现有表达和表型数据，通过分析UDN患者的人类表型本体注释并利用斑马鱼核心与斑马鱼信息网络(ZFIN)的连接进行优先排序。然后，斑马鱼核心将利用CRISPR/Cas9收集现有突变或产生新的突变，并执行高通量表型分析。拟议的MOSC还计划每两个月与统一数字网络站点和中心共享数据，并建立一个可供整个统一数字网络访问的网站，显示成果和正在进行的工作。不会提到个人信息、患者信息或症状，我们只会与直接参与该项目的调查人员分享。我们还概述了领导层与医生互动以获取诊断或医学上可操作的数据的计划。因此，拟建的中心使用了人类基因组学中最具创新性的技术--果蝇和斑马鱼，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.