Functional genomics resources for the Drosophila - TR&D3

项目概要- TR&D3 在果蝇系统中用于体内的蛋白质技术的发展已经落后于发展 分子遗传学技术。在这个项目中，我们将开发基于蛋白质的技术， 并且扩展了在该示例性模型系统中可以进行的研究的类型。我们将专注于发展 近端标记，如过氧化物酶或生物素连接酶促进的，作为体内蛋白质组学分析的工具， 特定的亚细胞区室和信号传导。我们还将开发一个强大的管道， 纳米抗体可用于多种应用，包括用于体内表达融合蛋白， 便于可视化和功能性阻断。对使用技术进行研究感兴趣的合作者 细胞生物学、发育、转录控制和神经元网络提供了适当的检测方法， 测试和改进。总之，这个项目将有助于填补果蝇工具箱中的一个重要空白， 可以帮助提供亚细胞、细胞、器官和整个动物的功能的更完整的图片 这些水平在细胞系统中或仅使用分子遗传工具无法实现。

PROJECT SUMMARY – TR&D3 Development of protein technologies for use in vivo in the Drosophila system has lagged behind development of molecular genetic technologies. In this project, we will develop protein-based technologies that complement and extend the type of studies that can be done in this exemplary model system. We will focus on development of proximity labeling, as facilitated by peroxidases or biotin ligases, as tools for in vivo proteomics analysis of specific subcellular compartments and signaling. We will additionally develop a robust pipeline for isolation of nanobodies that can be used in diverse applications, including for in vivo expression of fusion proteins that facilitate visualization and functional blocking. Collaborators with interests in using the technologies to study cell biology, development, transcription control, and neuronal networks provide appropriate assays for iterative testing and improvement. Altogether, this project will help fill an important gap in the Drosophila toolbox that can help provide a more complete picture of functions at the subcellular, cellular, organ, and whole animal levels that would not be achievable in cell systems or using only molecular genetic tools.