Godwin K Dogbevia^1,2, Martin Roβmanith¹, Rolf Sprengel³ and Mazahir T Hasan^1,4

Molecular Therapy Nucleic Acids (2016) 5, e309; doi:10.1038/mtna.2016.23
 

Abstract

Controlling gene expression in mammalian brain is of utmost importance to causally link the role of gene function to cell circuit dynamics under normal conditions and disease states. We have developed recombinant adeno-associated viruses equipped with tetracycline-controlled genetic switches for inducible and reversible control of gene expression in a cell type specific and brain subregion selective manner. Here, we characterize a two-virus approach to efficiently and reliably switch gene expression on and off, repetitively, both in vitro and in vivo. Our recombinant adeno-associated virus (rAAV)-Tet approach is highly flexible and it has great potential for application in basic and biomedical neuroscience research and gene therap

http://www.nature.com/mtna/journal/v5/n4/full/mtna201623a.html

Fig. 2: Functional gene expression in vitro and in vivo. (a) Dox-controlled, rtTA and tTA dependent gene activation in rat organotypic hippocampal slices infected with two viruses as depicted in the images above (scale bar 500 µm). With the rtTA system, tdTOM expression was undetectable in the absence of Dox (left panel, but it was robust with Dox (middle panel). With the tTA system, however, there was very strong tdTOM expression in the absence of Dox (right panel). (b) Firefly luciferase activity measurements of lysates from infected organotypic hippocampal slices treated with or without Dox. The mean of the luciferase activity in the absence of Dox was equated to 1. The data shown are mean of 4 independent infected organotypic slices and expressed as relative light units (RLU; fLUC/rLUC ± SEM). (c) In vivo bioluminescence in the somatosensory cortex in two mice infected with eLUC gene under control of a human hSYN promoter. Bioluminescence signal was detectable only after intraperitoneal injection of D-luciferin.