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A versatile genetic tool for post-translational control of gene expression in Drosophila melanogaster
eLife ( IF 6.4 ) Pub Date : 2017-11-15 , DOI: 10.7554/elife.30327
Sachin Sethi 1 , Jing W Wang 1
Affiliation  

Several techniques have been developed to manipulate gene expression temporally in intact neural circuits. However, the applicability of current tools developed for in vivo studies in Drosophila is limited by their incompatibility with existing GAL4 lines and side effects on physiology and behavior. To circumvent these limitations, we adopted a strategy to reversibly regulate protein degradation with a small molecule by using a destabilizing domain (DD). We show that this system is effective across different tissues and developmental stages. We further show that this system can be used to control in vivo gene expression levels with low background, large dynamic range, and in a reversible manner without detectable side effects on the lifespan or behavior of the animal. Additionally, we engineered tools for chemically controlling gene expression (GAL80-DD) and recombination (FLP-DD). We demonstrate the applicability of this technology in manipulating neuronal activity and for high-efficiency sparse labeling of neuronal populations.



中文翻译:

用于果蝇果蝇基因表达的翻译后控制的通用遗传工具

已经开发了几种技术来在完整的神经回路中暂时操纵基因表达。但是,目前开发用于果蝇体内研究的工具的适用性受到其与现有GAL4品系不兼容以及对生理和行为的副作用的限制。为了避免这些局限性,我们采用了一种策略,即通过使用去稳定域(DD)来可逆地调节小分子蛋白质的降解。我们表明,该系统在不同的组织和发育阶段均有效。我们进一步表明,该系统可用于体内控制基因表达水平低,背景动态范围大,并且可逆,对动物的寿命或行为无可检测的副作用。此外,我们设计了用于化学控制基因表达(GAL80-DD)和重组(FLP-DD)的工具。我们证明了该技术在操纵神经元活动和高效稀疏标记神经元人群中的适用性。

更新日期:2017-11-16
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