Gene Targeting in Drosophila

描述(由申请人提供)：基因组计划的最终目标必须 是为了了解基因的作用，而不仅仅是识别它们的存在。 我们在果蝇身上实现这一目标还有一段路要走。这个 基因打靶方法的可用性只是迈向这一目标的第一步 进球。最终，希望每个基因都有一个基因敲除突变。 基因能够解释这13,600个基因中的每一个的功能 基因。已经开发的目标定位方法要高效得多。 而且比之前为苍蝇开发的方法更有效，但 这种方法还处于初级阶段。系统的许多参数都是未知的， 目前这是一个漫长的过程，需要大约4-6个月的时间 从靶向结构的设计到击倒的生产 突变。拟议的工作旨在增加我们对 影响方法效率的参数，导致更高效 苍蝇的结构设计和操纵。制作的方法多种多样 将对基因敲除进行测试和表征，以找到那些 有效的，通常适合于将突变引入特定的基因。 还将进行实验，以开发加速这一过程的方法。 进行突变，需要几周甚至几个月。这项工作的结果将是 提供关于生产特定产品的最有效途径的指南 果蝇基因组的改变。此外，结果将决定 考虑在中国进行大规模淘汰赛项目是否可行 果蝇，以及这样一个项目的时间尺度是什么。最后， 果蝇基因打靶的有效方法将有助于 利用果蝇作为理解人类疾病生物学的模型。

DESCRIPTION (provided by applicant): The ultimate goal of a genome project must be to understand what the genes are doing, not merely identify their existence. We are yet some distance from achieving that goal in Drosophila. The availability of a gene targeting method is but the first step towards that goal. Ultimately, it would be desirable to have a knockout mutation for every gene in the fly to be able to elucidate the function of each of those 13,600 genes. The targeting method that has been developed is vastly more efficient and effective than prior methods that have been developed for flies, but the method is in its infancy. Many parameters of the system are uncharacterized, and it is currently a lengthy procedure, requiring approximately 4-6 months from design of the targeting construct to the production of a knockout mutation. The proposed work is designed to increase our knowledge of the parameters that affect the efficiency of the method, leading to more efficient construct design and manipulation of the flies. A variety of methods to make gene knockouts will be tested and characterized to find those that are efficient and generally suitable for introducing mutations into specific genes. Experiments will also be undertaken to develop methods to speed up the process of making mutations, by weeks or perhaps months. The result of this work will provide a guide to the most efficient route towards producing specific alterations in the Drosophila genome. Moreover, the results will determine whether it becomes feasible to contemplate a large-scale knockout project in Drosophila, and what the time scale of such a project will be. Finally, the availability of efficient methods for gene targeting in Drosophila will aid the use of Drosophila as a model for understanding the biology of human diseases.