Development of Validated Drosophila in vivo RNAi Models of Human Diseases

描述（由申请人提供）：果蝇与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与苍蝇有可识别的同源基因，而且所有主要的信号转导途径在苍蝇和人类之间都是保守的。RNAi可用于基因特异性敲低mRNA水平，这加快了我们进行分子遗传分析的步伐，正是这种分析使果蝇成为如此强大的系统。然而，尽管目前可用的工具令人印象深刻，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，无法获得具有良好特征的功能缺失等位基因或经过验证的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的基础上，以产生大量的体内果蝇RNAi人类疾病模型，即HuDis-TRiP疾病模型资源。这个资源将是独一无二的，其规模庞大，针对大约900个人类疾病基因的果蝇同源物，并在每个RNAi苍蝇种群中应用高水平的质量控制。对于基因功能丧失与疾病状态相关的人类疾病基因，HuDis-TRiP蝇群可能通过在细胞、组织和/或生物体水平上模拟人类疾病状态而充当疾病模型。具体而言，我们建议：目标1，利用在线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；目的2，鉴定900个高置信度苍蝇同源基因；每个基因制造两个转基因果蝇RNAi蝇群；进行表型鉴定、定量PCR和抢救以验证资源；和目标3，进一步描述最有前途的模型。值得注意的是，我们最初的保守疾病相关基因列表包括与几乎所有NIH研究所相关的基因。为了履行我们以社区为基础的努力承诺，所有RNAi蝇类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。我们预计这一资源将被社区广泛使用，因为它将使科学家能够在一个强大的遗传系统中立即开始高质量的、经过验证的疾病模型的研究。

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.