title: Polyswitch lentivectors: "all-in-one" lentiviral vectors for drug-inducible gene expression, live selection, and recombination cloning.
abstract: Lentiviral vectors are now widely considered one of the safest and most efficient tools for gene delivery and stable gene expression. Even though inducible gene expression cassettes are mandatory for many genetic engineering strategies, most current systems suffer from various issues, such as the requirement of two vectors, which decreases the overall efficiency of the transduction, leakiness and/or insufficient levels of activation of the inducible promoter, lack of selectable marker, low titers, or general issues associated with the cloning of large plasmids. In this article, we describe the design and functional characterization of a set of "all-in-one" multicistronic autoinducible lentivectors. They combine: (1) an optimized drug-inducible promoter; (2) a multicistronic strategy to express living color, selectable marker, and transactivator; and (3) acceptor sites for easy recombination cloning of genes of interest. These polyswitch lentivectors have good titers, very low basal activity, and reversible high induced activity, and can accept a growing number of genes already cloned in entry plasmids. These combined features make them a novel, powerful, and versatile tool for current and future genetic engineering approaches.
authors.affiliation: Department of Neurosciences, Faculty of Medicine, CMU, 1 Rue Michel Servet, Geneva, Switzerland.
authors.forename: Marc
authors.initials: M
authors.lastname: Giry-Laterrière
authors.forename: Ophélie
authors.initials: O
authors.lastname: Cherpin
authors.forename: Yong-Sik
authors.initials: YS
authors.lastname: Kim
authors.forename: Jan
authors.initials: J
authors.lastname: Jensen
authors.forename: Patrick
authors.initials: P
authors.lastname: Salmon
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D015153:Blotting, Western; D017931:DNA Primers / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics*; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D016086:Lentivirus; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D022821:RNA Splice Sites / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; C068356:CcdB protein, Plasmid F; D017931:DNA Primers; D022821:RNA Splice Sites; D015534:Trans-Activators