title: Generation and evaluation of an IPTG-regulated version of Vav-gene promoter for mouse transgenesis.
abstract: Different bacteria-derived systems for regulatable gene expression have been developed for the use in mammalian cells and some were also successfully adopted for in vivo use in vertebrate model organisms. However, certain limitations apply to most of these systems, including leakiness of transgene expression, inefficient transgene silencing or activation, as well as limited tissue accessibility of transgene-inducers or their unfavourable pharmacokinetics. In this study, we evaluated the suitability of the lac-operon/lac-repressor (lacO/lacI) system for the regulation of the well-established Vav-gene promoter that allows inducible transgene expression in different haematopoietic lineages in mice. Using the fluorescence marker protein Venus as a reporter, we observed that the lacO/lacI system could be amended to modulate transgene-expression in haematopoietic cells. However, reporter expression was not uniform and the lacO elements introduced into the Vav-gene promoter only conferred limited repression and reversion of lacI-mediated gene silencing after administration of IPTG. Although further optimization of the system is required, the lacO-modified version of the Vav-gene promoter may be adopted as a tool where low basal gene-expression and limited transient induction of protein expression are desired, e.g. for the activation of oncogenes or transgenes that act in a dominant-negative manner.
authors: Division of Developmental Immunology, Medical University Innsbruck, Innsbruck, Austria.
authors: Francesca
authors: F
authors: Grespi
authors: Eleonora
authors: E
authors: Ottina
authors: Nikolaos
authors: N
authors: Yannoutsos
authors: Stephan
authors: S
authors: Geley
authors: Andreas
authors: A
authors: Villunger
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA; D005786:Gene Expression Regulation / Q000187:drug effects*; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology*; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D051567:Proto-Oncogene Proteins c-vav / Q000235:genetics*
chemical_list: D051567:Proto-Oncogene Proteins c-vav; D007544:Isopropyl Thiogalactoside; D004247:DNA