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通过组合过表达和抑制策略,在果蝇翅膀的内源性表达区域中灵活操纵 Omb 水平。

Flexible manipulation of Omb levels in the endogenous expression region of Drosophila wing by combinational overexpression and suppression strategy.

机构信息

Institute of Applied Biology, College of Life Science, Shanxi University, Taiyuan, China.

Applied Zoology, Technical University Dresden, Zellescher Weg 20b, Dresden, Germany.

出版信息

Insect Sci. 2020 Feb;27(1):14-21. doi: 10.1111/1744-7917.12705. Epub 2019 Jul 23.

Abstract

Manipulating an exogenous or endogenous gene of interest at a defined level is critical for a wide variety of experiments. The Gal4/UAS system has been widely used to direct gene expression for studying complex genetic and biological problems in Drosophila melanogaster and other model organisms. Driven by a given tissue-specific Gal4, expressing UAS-transgene or UAS-RNAi (RNA interference) could be used to up- or down-regulate target gene expression, respectively. However, the efficiency of the Gal4/UAS system is roughly predefined by properties of transposon vector constructs and the insertion site in the transgenic stock. Here, we describe a simple way to modulate optomotor blind (omb) expression levels in its endogenous expression region of the wing disc. We co-expressed UAS-omb and UAS-omb-RNAi together under the control of dpp-Gal4 driver which is expressed in the omb expression region of the wing pouch. The repression effect is more sensitive to temperature than that of overexpression. At low temperature, overexpression plays a dominant role but the efficiency is attenuated by UAS-omb-RNAi. In contrast, at high temperature RNAi predominates in gene expression regulation. By this strategy, we could manipulate omb expression levels at a moderate level. It allows us to manipulate omb expression levels in the same tissue between overexpression and repression at different stages by temperature control.

摘要

在特定水平上操纵感兴趣的外源或内源性基因对于各种实验都至关重要。Gal4/UAS 系统已被广泛用于指导基因表达,以研究黑腹果蝇和其他模式生物中的复杂遗传和生物学问题。在特定组织的 Gal4 驱动下,表达 UAS 转基因或 UAS-RNAi(RNA 干扰)可分别上调或下调靶基因的表达。然而,Gal4/UAS 系统的效率大致由转座子载体构建体的特性和转基因品系中的插入位点预先确定。在这里,我们描述了一种在翅膀盘的 omb 表达区域内调节视动盲(omb)表达水平的简单方法。我们共同表达 UAS-omb 和 UAS-omb-RNAi,受 dpp-Gal4 驱动控制,该驱动在翅膀囊的 omb 表达区域表达。抑制效应对温度比过表达更敏感。在低温下,过表达起主导作用,但 UAS-omb-RNAi 会降低效率。相比之下,在高温下,RNAi 在基因表达调控中占主导地位。通过这种策略,我们可以将 omb 的表达水平调节到中等水平。它允许我们通过温度控制在不同阶段在同一组织中操纵 omb 的表达水平,使其在过表达和抑制之间进行转换。

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