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: 10676847
• 负责人: Ying Tan
• 金额: $ 83.71万
• 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/10676847
• 关键词: Acceleration; 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; Letters; Life Cycle Stages; Mammals; Marketing; Mediating; Methods; Mitosis; Molecular; 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 conservation; gene function; genetic manipulation; human disease; insight; loss of function; loss of function mutation; mosaic analysis; 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.

项目总结/摘要 该提案的总体目标是生成、维护和分发一个新的2，800个条件 超过1,400个果蝇基因的功能丧失和获得等位基因， 在人类中保存。使果蝇区别于其他模型系统的一个关键因素是巨大的 在过去100年的研究中积累了丰富的遗传和分子工具。与 一个庞大而活跃的果蝇研究社区，额外的技术和工具，使整个领域受益 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一种 动物模型系统由于果蝇和哺乳动物在基因表达方面存在许多相似之处， 控制生物过程的潜在分子机制，从研究中获得的知识， 果蝇既可以直接应用，也可以很容易地适应人类生物学和疾病的理解。 我们建议产生一组条件性的“flip-flop-loxP”等位基因，这些等位基因允许特定的基因被 在果蝇的任何细胞类型中，大部分保守基因在任何时候都是关闭或打开的。 因此，所产生的集合与现有资源不同，因为它允许不依赖于有丝分裂的 基因活性的调节，使得能够在不同发育阶段进行基因功能的镶嵌分析 在成年人中也是如此。此外，该方法允许恢复具有全细胞类型和时间的基因功能。 控制因此，这一新的收集将使几乎所有的果蝇研究人员受益;因此， 在今后许多年里，对这种资源的需求将很大。我们建议产生这样的等位基因 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个功能丧失和功能获得的触发器loxP等位基因。 目标二。创建并维护一个可在线搜索的数据库，用于收集触发器loxP。 这种条件等位基因的集合将通过增加显著的能力来补充现有的资源， 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。一旦生成，这个集合将为果蝇提供很大的实用性， 研究社区。作为潜在市场规模巨大的明显证据， 全世界的果蝇实验室。此外，目前仅NIH资助的项目就有2,567个 在资助的标题中有“果蝇”，包括每年超过11亿美元的总费用。 由于许多使用果蝇作为模型系统的赠款在标题中不包括属名，这些 这些数字显然低估了潜在的市场规模。因此，有一个大的和积极的资助， 研究社区，代表了广泛实用的新试剂的巨大市场。