Gene Targeting in Drosophila

描述（由申请人提供）：基因组计划的最终目标必须 我们需要了解基因在做什么，而不仅仅是确定它们的存在。 我们离在果蝇中实现这一目标还有一段距离。的 基因靶向方法的可用性只是实现这一目标的第一步， 目标.最后，我们希望有一个敲除突变， 果蝇中的基因能够阐明这13600个基因中每一个的功能 基因.已经开发出的瞄准方法要有效得多 和有效的比以前的方法已经开发的苍蝇，但 方法尚处于起步阶段。系统的许多参数是未表征的， 目前这是一个漫长的过程，需要大约4-6个月 从靶向构建体的设计到敲除的产生 突变拟议的工作旨在增加我们对 影响方法效率的参数，导致更有效的 构建设计和操纵苍蝇。多种方法使 基因敲除将被测试和表征，以找到那些 有效且通常适合于将突变引入特定基因。 还将进行实验，以制定加快这一进程的方法 几周甚至几个月的时间来制造变异。这项工作的结果将 提供了一个指南，以最有效的途径， 果蝇基因组的改变此外，结果将决定 考虑一个大规模的淘汰项目是否可行， 以及这个项目的时间范围。最后 果蝇基因打靶的有效方法的可用性将有助于 使用果蝇作为了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.