A Conditional Loss-of-Function Mutation Collection in Drosophila

果蝇条件性功能丧失突变集合

• 批准号: 10546055
• 负责人: Ying Tan
• 金额: $ 89.36万
• 依托单位: GENETIVISION CORPORATION
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546055
• 关键词: Address; Adult; Alleles; Animal Model; Applied Research; Basic Science; Biological Models; Biological Process; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collaborations; Collection; Communities; Complement; Custom; DNA cassette; Development; Drosophila genus; Drosophila melanogaster; Exons; FlyBase; Funding; Gene Silencing; Genes; Genetic; Genetic Recombination; Goals; Grant; Human; Human Biology; Human Development; Introns; Knock-out; Knowledge; Laboratories; Lead; Letters; Life Cycle Stages; Mammals; Mediating; Methods; Mitosis; Molecular; Mosaicism; Names; Phase; Production; Reagent; Reporter; Research; Research Personnel; Resources; Site; Small Business Innovation Research Grant; Source; Technology; Testing; Time; Tissues; Transgenic Organisms; United States National Institutes of Health; Update; cell type; cost; design; flexibility; fly; gain of function; gene function; genetic manipulation; human disease; insight; loss of function mutation; recombinase-mediated cassette exchange; relational database; restoration; searchable database; tool; virtual; web site

Project Summary/Abstract The overall goal of this proposal is to generate, maintain, and distribute a new collection of 2,800 conditional loss- and gain-of-function alleles for more than 1,400 Drosophila melanogaster genes that are highly conserved in humans. One key factor that sets Drosophila apart from other model systems is the huge wealth of genetic and molecular tools that have accumulated in the past 100 years of research. With a large, active Drosophila research community, additional technologies and tools that benefit the entire field can have a profound impact by accelerating the pace of research for many. Many of the most important advances in our understanding of human development have come from studies using the fruit fly as an animal model system. Since many parallels exist between Drosophila and mammals in terms of the underlying molecular mechanisms controlling biological processes, knowledge gained from research in Drosophila can be either directly applied or readily adapted to understanding human biology and disease. We propose to generate a collection of conditional "flip-flop-loxP" alleles that allow a specific gene to be turned off or on at any time in any cell type for a large fraction of the conserved genes in Drosophila. Therefore, the resulting collection is distinct from existing resources as it allows mitosis-independent modulation of gene activity, enabling mosaic analysis of gene function during different development stages and in adults. Moreover, this method allows restoration of gene function with full cell-type and temporal control. Therefore, this new collection will benefit virtually all Drosophila researchers; as such it is likely that there will be a high demand for this resource for many years to come. We propose to produce such alleles for 1,400 highly conserved genes. Our Specific Aims are to: Aim 1. Generate and validate 2,800 loss- and gain-of-function flip-flop-loxP alleles. Aim 2. Create and maintain an online searchable database for the flip-flop-loxP collection. This collection of conditional alleles will complement existing resources by adding significant capability to investigate gene function in any tissue/cell type in developing or adult flies by either knockout or restoration of function at any desired time. Once generated, this collection will offer great utility for the Drosophila research community. As clear evidence of the large size of the potential market, there are at least 2,000 Drosophila laboratories worldwide. Moreover, there are currently 2,567 projects funded by the NIH alone that have "Drosophila" in the title of the grant, comprising more than 1.1 billion USD in total costs per year. Since many grants that use Drosophila as a model system do not include the genus name in the title, these numbers are a clear underestimation of the potential market size. Thus, there is a large and actively funded research community that represents a substantial market for new reagents of broad utility.

项目概要/摘要 该提案的总体目标是生成、维护和分发包含 2,800 个条件的新集合 超过 1,400 个果蝇基因的功能丧失和获得等位基因，这些基因高度 保存在人类体内。果蝇与其他模型系统区别开来的一个关键因素是巨大的 过去 100 年研究中积累的丰富的遗传和分子工具。与一个 大型、活跃的果蝇研究社区，有利于整个领域的其他技术和工具 可以通过加快许多人的研究步伐来产生深远的影响。许多最重要的 我们对人类发展的理解的进步来自于使用果蝇作为研究对象的研究 动物模型系统。由于果蝇和哺乳动物之间存在许多相似之处 控制生物过程的潜在分子机制，从研究中获得的知识 果蝇可以直接应用或很容易适应理解人类生物学和疾病。 我们建议生成条件“flip-flop-loxP”等位基因的集合，这些等位基因允许特定基因 果蝇中大部分保守基因在任何细胞类型中都可以随时关闭或打开。 因此，所得的集合与现有资源不同，因为它允许有丝分裂独立 调节基因活性，实现不同发育阶段基因功能的镶嵌分析 和成人。此外，该方法允许恢复全细胞类型和时间的基因功能 控制。因此，这个新集合几乎将使所有果蝇研究人员受益；因此很可能 未来许多年对这一资源的需求将会很高。我们建议生产这样的等位基因 1,400 个高度保守的基因。我们的具体目标是： 目标 1. 生成并验证 2,800 个功能丧失和功能增强的触发器-loxP 等位基因。 目标 2. 创建并维护 Flip-flop-loxP 集合的在线可搜索数据库。 这个条件等位基因的集合将通过增加显着的能力来补充现有资源 通过敲除或恢复研究发育中或成年果蝇中任何组织/细胞类型的基因功能 在任何需要的时间发挥作用。一旦生成，该集合将为果蝇提供巨大的实用性 研究社区。作为潜在市场规模之大的明确证据，至少有 2,000 世界各地的果蝇实验室。此外，目前仅 NIH 资助的项目就有 2,567 个 资助名称中含有“果蝇”的项目，每年总成本超过 11 亿美元。 由于许多使用果蝇作为模型系统的资助在标题中不包括属名，这些 数字明显低估了潜在市场规模。因此，有一个庞大且积极资助的 研究界代表了具有广泛用途的新试剂的巨大市场。