Development of Validated Drosophila in vivo RNAi Models of Human Diseases

开发经过验证的果蝇体内人类疾病 RNAi 模型

• 批准号: 8327181
• 负责人: NORBERT PERRIMON
• 金额: $ 43.99万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8327181
• 关键词: Affect; Aging; Alleles; Animal Model; Behavior; Biological Assay; Biological Models; Biological Process; Brain; Cells; Collection; Communities; Data; Data Set; Databases; Defect; Development; Disease; Disease model; Drosophila genus; Eye; Fertility; Gene Expression Profile; Genes; Genetic; Human; Infection; Institutes; Laboratories; Locomotion; Memory; Messenger RNA; Metabolism; Modeling; Molecular; Molecular Genetics; Morphology; Online Mendelian Inheritance In Man; Organ; Organism; Orthologous Gene; Pathway interactions; Pharmaceutical Preparations; Phenotype; Preparation; Process; Quality Control; RNA Interference; Research Infrastructure; Research Personnel; Resources; Scientist; Signal Transduction Pathway; Source; Staging; System; Testing; Time; Tissues; Transgenic Organisms; United States National Institutes of Health; Wing; Work; assay development; base; cost effective; design; fly; follow-up; gain of function mutation; gene function; genetic analysis; human disease; improved; in vivo; innovation; insight; interest; loss of function; malignant neurologic neoplasms; nervous system disorder; research study; small hairpin RNA; tool; vector; web site

DESCRIPTION (provided by applicant): Remarkable conservation exists between Drosophila and humans at the level of genes and gene functions. For example, there are identifiable fly orthologs for about two-thirds of human disease genes and all of the major signal transduction pathways have been conserved between flies and humans. The availability of RNAi for gene-specific knockdown of mRNA levels has accelerated the pace at which we can undertake the type of molecular genetic analyses that make Drosophila such a powerful system. However, despite the impressive set of tools currently available, locating and/or generating Drosophila disease models can be time consuming, and in most cases, well-characterized loss-of-function alleles or validated RNAi strains are not available. The proposed project would build upon our existing expertise and infrastructure to produce a large collection of in vivo Drosophila RNAi models of human disease, the HuDis-TRiP Resource of Disease Models. This resource will be unique in its large scale, targeting approximately 900 Drosophila orthologs of human disease genes, and in the high level of quality control applied to each RNAi fly stock. For human disease genes for which loss-of-function of the gene is associated with disease state, HuDis-TRiP fly stocks are likely to serve as disease models by mimicking the human disease state at the cell, tissue and/or organism level. Specifically, we propose to: Aim 1, compile a prioritized list of human disease genes using the Online Mendelian Inheritance in Man database and a community nomination process; Aim 2, identify 900 high-confidence fly orthologs of these genes; make two transgenic Drosophila RNAi fly stocks per gene; perform phenotypic characterization, quantitative PCR and rescue to validate the resource; and Aim 3, further characterize the most promising models. Notably, our initial list of conserved disease-associated genes includes genes relevant to nearly all NIH Institutes. In keeping with our commitment to community-based efforts, all RNAi fly stocks will be rapidly transferred to a public stock center and all datasets will be made available through our own database, website (www.flyrnai.org) and other databases. We anticipate that this resource will be widely used by the community, as it will allow scientists to immediately begin their studies with high quality, validated disease models in a powerful genetic system.

描述（申请人提供）：果蝇和人类在基因和基因功能水平上存在显著的保守性。例如，大约三分之二的人类疾病基因存在可识别的果蝇直系同源物，并且所有主要的信号转导途径在果蝇和人类之间都是保守的。RNA干扰技术在基因特异性敲低mRNA水平上的应用加快了我们进行分子遗传学分析的步伐，使果蝇成为一个如此强大的系统。然而，尽管目前有一套令人印象深刻的工具，但定位和/或生成果蝇疾病模型可能非常耗时，并且在大多数情况下，无法获得充分表征的功能丧失等位基因或验证的RNAi菌株。拟议的项目将建立在我们现有的专业知识和基础设施，以产生大量的人类疾病的体内果蝇RNAi模型，HuDis-TRiP疾病模型资源。这一资源将是独一无二的，其规模大，针对约900个人类疾病基因的果蝇直系同源物，并在高水平的质量控制应用于每一个RNAi苍蝇股票。对于基因功能丧失与疾病状态相关的人类疾病基因，HuDis-TRiP蝇种可能通过在细胞、组织和/或生物体水平上模拟人类疾病状态而用作疾病模型。具体而言，我们建议：目标1，使用在线人类孟德尔遗传数据库和社区提名程序编制人类疾病基因的优先列表;目标2，确定这些基因的900个高置信度的果蝇直系同源物;每个基因制作两个转基因果蝇RNAi果蝇种群;进行表型表征，定量PCR和拯救以验证资源;目标3，进一步表征最有希望的模型。值得注意的是，我们最初的保守疾病相关基因列表包括几乎所有NIH研究所相关的基因。为了与我们对社区工作的承诺保持一致，所有RNAi苍蝇库存将迅速转移到公共库存中心，所有数据集将通过我们自己的数据库，网站（www.flyrnai.org）和其他数据库提供。我们预计该资源将被社区广泛使用，因为它将使科学家能够立即开始在强大的遗传系统中使用高质量、经验证的疾病模型进行研究。