A versatile system for cell-specific control of gene expression in the fly brain

描述(由申请人提供)：现代神经科学寻求识别并精确操作大脑中特定的细胞类型，以识别电路组装、功能和神经疾病的机制。这些技术是从系统神经科学到分子发育分析等大量研究领域的先决条件，不仅对神经功能，而且对包括衰老、神经退化和神经修复在内的疾病过程提供见解的关键。开发工具来标记和操作特定的神经元类型通常是昂贵和耗时的。这个应用程序的目标是开发并向科学界发布一个广泛而通用的工具包，用于操纵果蝇的神经系统，这是一个强大的遗传模型，已经产生了对神经系统功能和发育的许多基本见解。这个应用程序有三个中心目标。首先，我们建议在最近开发的INSITE系统的基础上创造4000个新的转基因株系。这些菌株将允许对果蝇绝大多数神经元中的基因表达进行细胞特异性操作。这些插入位点中的每一个都将被分子作图，每一条线捕捉到的表达模式将在幼虫和成虫的大脑中进行描述。其次，使用这个集合，我们将获得细胞类型特定的交叉工具，以及独立的转录调控系统，允许在多种单独的细胞类型中独立地操纵基因表达。第三，我们将创建一个描述这些数据的数据库，使社区能够充分利用这些资源来解决广泛的科学问题。这个集合将提供一个平台，将基因表达的标记和驱动因素的多功能集合引入存在于幼虫和成年神经系统中的大量神经元类型，以便释放到果蝇群落。一旦建立了这个资源，使用和扩大收集的方法只涉及可以在任何实验室中适应的基本遗传杂交。我们期望，这个集合将为神经系统操作和基因发现提供一个不断扩大的、高度通用的、具有成本效益的工具集合。

DESCRIPTION (provided by applicant): Modern neuroscience seeks to identify, and manipulate with precision, specific cell types in the brain to identify mechanisms of circuit assembly, function, and neural disease. These technologies are a prerequisite to a large number of fields of study, from systems neuroscience to molecular developmental analysis, and are key to providing insights not only into neural function, but also disease processes, including aging, neurodegeneration, and neural repair developing tools to label and manipulate specific neuronal types is generally costly and time-consuming. The objective of this application is to develop and release to the scientific community an extensive and versatile toolkit for manipulation of the nervous system of Drosophila, a powerful genetic model that has generated many fundamental insights into nervous system function and development. This application has three central aims. First, we propose to generate 4,000 new transgenic lines based on the recently developed InSITE system. These strains will allow cell-specific manipulations of gene expression in the vast majority of neurons in the fly. Each of these insertion sites will be molecularly mapped, and the expression pattern captured by each line will be described in both larval and adult fly brains. Second, using this collection, we will derive cell-type specific intersectional tools, as well as independent transcriptional regulatory systems, allowing gene expression to be manipulated in multiple individual cell types, independently. Third, we will create a database describing these data that will enable the community to make full use of this resource to address a wide range of scientific questions. This collection will provide a platform for introducing a versatile collection of markers and drivers of gene expression into a large number of the neuronal types present in the larval and adult nervous systems for release to the Drosophila community. Once this resource is established, the methods for using and extending the collection involve only basic genetic crosses that can be adapted in any lab. We expect that this collection will seed an expanding, highly versatile, and cost-effective collection of tools fo manipulation of the nervous system and gene discovery.