title: Scoring and manipulating gene position and dynamics using FROS in budding yeast.
abstract: The spatial organization of the genome within the nucleus is now seen as a key contributor to genome function. Studying chromatin dynamics in living cells has been rendered possible by the development of fast microscopy coupled with fluorescent repressor operator systems (FROS). In these systems, arrays of protein-binding sites integrated at specific loci by homologous recombination are monitored through the fluorescence of tagged DNA-binding proteins. In the budding yeast, where homologous recombination is efficient, this technique, combined with targeting assay and genetic analysis, has been extremely powerful for studying the determinants and function of chromatin dynamics in living cells. However, issues have been recurrently raised in different species regarding the use of these systems. Here we discuss the different uses of gene tagging with FROS and their limitations, focusing in budding yeast as a model organism.
authors.affiliation: Institut Curie, Centre de Recherche, Paris, France.|Centre National de la Recherche Scientifique (CNRS), UMR 3364, Paris, France.|Université Pierre-et-Marie-Curie (UPMC), UMR 3664, Paris, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Loïodice
authors.affiliation: Université Pierre-et-Marie-Curie (UPMC), UMR 3664, Paris, France.|Institute for Cell and Molecular Biosciences, Medical School, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Marion
authors.initials: M
authors.lastname: Dubarry
authors.affiliation: Institut Curie, Centre de Recherche, Paris, France.|Centre National de la Recherche Scientifique (CNRS), UMR 3364, Paris, France.|Université Pierre-et-Marie-Curie (UPMC), UMR 3664, Paris, France.
authors.forename: Angela
authors.initials: A
authors.lastname: Taddei
mesh_terms: D002843:Chromatin / Q000235:genetics* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D004718:Saccharomycetales / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*
chemical_list: D002843:Chromatin; D049452:Green Fluorescent Proteins
keywords: FROS; Saccharomyses cerevisiae; lacO; nuclear organization; tetO