A versatile genetic tool for post-translational control of gene expression in Drosophila melanogaster.

Jing W Wang,Sachin Sethi

doi:10.7554/elife.30327

Abstract

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.

Highlights

Tools for precise spatial and temporal control of gene expression are essential for understanding how neuronal circuits develop and function
We first tested whether the ecDHFR-derived destabilizing domain (DD) can be used to control GFP expression levels
TMPinduced DD stabilization is dose-dependent over several orders of magnitude of TMP concentration. This dose-dependency can be exploited for titration of in vivo gene expression levels

Summary

Introduction

Tools for precise spatial and temporal control of gene expression are essential for understanding how neuronal circuits develop and function. Drosophila melanogaster, bipartite expression systems (GAL4/UAS, LexA/LexAop, QF/QUAS) provide a powerful means to control gene expression in a spatially selective manner (Brand and Perrimon, 1993; Lai and Lee, 2006; Potter et al, 2010). Several modifications of these expression systems have been made to permit temporal control over the exogenous transcription factors (GAL4, LexA or QF) (Chan et al, 2015; McGuire et al, 2003; Osterwalder et al, 2001; Potter et al, 2010).

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