title: RANGE: Gene Transfer of Reversibly Controlled Polycistronic Genes.
abstract: We developed a single vector recombinant adeno-associated viral (rAAV) expression system for spatial and reversible control of polycistronic gene expression. Our approach (i) integrates the advantages of the tetracycline (Tet)-controlled transcriptional silencer tTS(Kid) and the self-cleaving 2A peptide bridge, (ii) combines essential regulatory components as an autoregulatory loop, (iii) simplifies the gene delivery scheme, and (iv) regulates multiple genes in a synchronized manner. Controlled by an upstream Tet-responsive element (TRE), both the ubiquitous chicken β-actin promoter (CAG) and the neuron-specific synapsin-1 promoter (Syn) could regulate expression of tTS(Kid) together with two 2A-linked reporter genes. Transduction in vitro exhibited maximally 50-fold regulation by doxycycline (Dox). Determined by gene delivery method as well as promoter, highly specific tissues were transduced in vivo. Bioluminescence imaging (BLI) visualized reversible "ON/OFF" gene switches over repeated "Doxy-Cycling" in living mice. Thus, the reversible rAAV-mediated N-cistronic gene expression system, termed RANGE, may serve as a versatile tool to achieve reversible polycistronic gene regulation for the study of gene function as well as gene therapy.Molecular Therapy - Nucleic Acids (2013) 2, e85; doi:10.1038/mtna.2013.15; published online 9 April 2013.
authors.affiliation: Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Heidelberg, Germany.
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Chen
authors.forename: Liji
authors.initials: L
authors.lastname: Cao
authors.forename: Chonglin
authors.initials: C
authors.lastname: Luo
authors.forename: Désirée Aw
authors.initials: DA
authors.lastname: Ditzel
authors.forename: Jörg
authors.initials: J
authors.lastname: Peter
authors.forename: Rolf
authors.initials: R
authors.lastname: Sprengel