A New Tool Kit for Whole-Genome Mapping and Complementation in Drosophila

描述（由申请人提供）：本提案的最终目标是生成、维护和分发一个新的复制种群集合，该集合涵盖了黑腹果蝇基因组的整个常染色体部分。由于这个集合将代表一套强大的新工具，这将使几乎所有果蝇遗传学家受益，因此很可能对该资源有很高的需求。在我们对人类发展的理解中，许多最重要的进展都来自于使用果蝇作为动物模型系统的研究。由于果蝇和哺乳动物在控制生物过程的潜在分子机制方面存在许多相似之处，因此从果蝇研究中获得的知识可以直接应用于或很容易适应于理解人类生物学。我们建议生成一个新的重复集合，它将填补当前分子定义果蝇缺失试剂盒（MDADK）中99.4%的碱基空缺，并将提供平均10倍的高分辨率。由于该复制试剂盒是基于产生携带基因组构建物的转基因果蝇，因此可以对相同构建物进行进一步操作以进行下游功能分析，包括蛋白质标记和结构-功能研究。我们估计需要为这个收集产生5,000到6,000个转基因种群，并将该项目分为两个阶段：第一阶段将作为原理证明并覆盖8%的常染色体基因组，而第二阶段将完成收集。我们将：1；选择和测序克隆以覆盖分子定义的常染色体缺陷试剂盒中的所有空白。产生、平衡和维持400个携带基因组重复的转基因种群，第一阶段的成功不仅填补了目前MDADK中99.4%的碱基缺口，而且为扩大我们的努力创造完整的收集奠定了基础。利用新技术的好处，这个集合将是果蝇研究工具包的另一个关键补充。作为世界上大多数果蝇遗传学家高度期望的资源，该集合将为整个果蝇研究界提供巨大的实用性，其应用范围包括遗传精细定位，基因组拯救，体细胞克隆分析和基因组标记。

DESCRIPTION (provided by applicant): The ultimate goal of this proposal is to generate, maintain, and distribute a new collection of duplication stocks that covers the entire euchromatic autosomal portion of the Drosophila melanogaster genome. Since this collection will represent a powerful new set of tools that will benefit virtually all Drosophila geneticists, it is likely that there will be a high demand for this resource. Many of the most important advances in our understanding of human development have come from studies using the fruit fly Drosophila melanogaster 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. We propose to generate a new duplication collection that will fill 99.4% of the bases in the gaps of the current molecularly defined Drosophila deletion kit (MDADK) and will offer an average of 10-fold higher resolution. As this duplication kit is based on generating transgenic flies carrying genomic constructs, further manipulation of the same constructs can be performed for downstream functional analysis, including tagging of proteins and structure-function studies. We estimate that between 5,000 and 6,000 transgenic stocks will need to be generated for this collection and will split this project into two phases: Phase I will serve as a proof of principle and cover 8% of the autosomal genome while Phase II will complete the collection. We will: 1. Select and sequence clones to cover all gaps in the molecularly defined autosomal deficiency kit 2. Generate, balance, and maintain 400 transgenic stocks carrying genomic duplications Success of Phase I will not only fill 99.4% of the bases in gaps in the current MDADK, but also set the stage for expanding our effort to generate the complete collection. Harnessing the benefits of new technologies, this collection will be another key addition to the Drosophila research tool kit. As a highly desired resource by most Drosophila geneticists across the world, this collection will offer great utility for the entire Drosophila research community, with applications ranging from genetic fine mapping, genomic rescue, somatic clonal analysis, and genomic tagging. PUBLIC HEALTH RELEVANCE: The main goal of this project is to create a new resource for the Drosophila (a fruit fly) research community that will allow rapid mapping and identification of new genes required for normal development and function. Specifically, we propose to create a large set of transgenic flies that will carry duplications that total approximately 80% of the Drosophila genome but in pieces that represent only a few genes each. Since the vast majority of genes in Drosophila are highly conserved in mammals and often provide critical insights into human gene function, this resource will greatly benefit both basic and applied research. Moreover, this resource will be in high demand for many years to come by the more than 1000 active Drosophila research laboratories around the world and will therefore create a highly marketable and profitable venture.