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Spatial and Temporal Control of Gene Expression in Drosophila Using the Inducible GeneSwitch GAL4 System. I. Screen for Larval Nervous System Drivers

机译:使用诱导型GeneSwitch GAL4系统在果蝇中基因表达的时空控制。 I.幼虫神经系统驱动程序的屏幕

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摘要

There is a critical need for genetic methods for the inducible expression of transgenes in specific cells during development. A promising approach for this is the GeneSwitch GAL4 system of Drosophila. With GeneSwitch GAL4 the expression of upstream activating sequence (UAS) effector lines is controlled by a chimeric GAL4 protein that becomes active in the presence of the steroid RU486 (mifepristone). To improve the utility of this expression system, we performed a large-scale enhancer-trap screen for insertions that yielded nervous system expression. A total of 204 GeneSwitch GAL4 lines with various larval expression patterns in neurons, glia, and/or muscle fibers were identified for chromosomes I–III. All of the retained lines show increased activity when induced with RU486. Many of the lines reveal novel patterns of sensory neurons, interneurons, and glia. There were some tissue-specific differences in background expression, with muscles and glia being more likely to show activity in the absence of the inducing agent. However, >90% of the neuron-specific driver lines showed little or no background activity, making them particularly useful for inducible expression studies.
机译:迫切需要用于在发育过程中在特定细胞中诱导转基因表达的遗传方法。一种有前途的方法是果蝇的GeneSwitch GAL4系统。使用GeneSwitch GAL4,上游激活序列(UAS)效应子系的表达由嵌合GAL4蛋白控制,该嵌合GAL4蛋白在类固醇RU486(米非司酮)存在下变得有活性。为了提高该表达系统的实用性,我们对产生神经系统表达的插入物进行了大规模的增强子捕获筛选。共鉴定了204条GeneSwitch GAL4品系,它们的I–III染色体在神经元,神经胶质和/或肌肉纤维中具有各种幼虫表达模式。当用RU486诱导时,所有保留的系显示出增加的活性。许多线条揭示了感觉神经元,中间神经元和神经胶质的新颖模式。在背景表达中存在一些组织特异性差异,在缺乏诱导剂的情况下,肌肉和神经胶质细胞更有可能表现出活性。但是,> 90%的神经元特异性驱动程序系几乎没有或没有背景活性,这使其特别适用于诱导表达研究。

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