An auxin-inducible, GAL4-compatible, gene expression system for Drosophila.
An auxin-inducible, GAL4-compatible, gene expression system for Drosophila.
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DOI：
10.7554/elife.67598
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发表时间：
2022-04-01
期刊：
影响因子：
7.7
通讯作者：
Southall TD
中科院分区：
文献类型：
--
作者：
McClure CD;Hassan A;Aughey GN;Butt K;Estacio-Gómez A;Duggal A;Ying Sia C;Barber AF;Southall TD
在空间和时间上控制转基因表达的能力对于研究模式生物是必不可少的。在果蝇中，空间控制主要由GAL 4/UAS系统提供，而时间控制依赖于温度敏感的GAL 80（抑制GAL 4）和药物诱导系统。然而，这些都不是理想的。温度变化会影响许多生理和行为特征，目前的药物诱导系统要么是泄漏的，有毒的，与现有的GAL 4驱动系不相容，要么不能产生有效的表达水平。在这里，我们描述了生长素诱导的基因表达系统（AGES）。AGES依赖于生长素依赖性降解一个普遍表达的GAL 80，因此，是与现有的GAL 4驱动线兼容。将水溶性生长素以低的非致死浓度添加到苍蝇食物中，其诱导的表达与未受抑制的GAL 4表达相当。该系统在幼虫和成虫中均起作用，为暂时控制果蝇中的GAL 4活性提供了一种严格的、非致死的、具有成本效益的和方便的方法。
The ability to control transgene expression, both spatially and temporally, is essential for studying model organisms. In Drosophila, spatial control is primarily provided by the GAL4/UAS system, whilst temporal control relies on a temperature-sensitive GAL80 (which inhibits GAL4) and drug-inducible systems. However, these are not ideal. Shifting temperature can impact on many physiological and behavioural traits, and the current drug-inducible systems are either leaky, toxic, incompatible with existing GAL4-driver lines, or do not generate effective levels of expression. Here, we describe the auxin-inducible gene expression system (AGES). AGES relies on the auxin-dependent degradation of a ubiquitously expressed GAL80, and therefore, is compatible with existing GAL4-driver lines. Water-soluble auxin is added to fly food at a low, non-lethal, concentration, which induces expression comparable to uninhibited GAL4 expression. The system works in both larvae and adults, providing a stringent, non-lethal, cost-effective, and convenient method for temporally controlling GAL4 activity in Drosophila.