title: A mobile insulator system to detect and disrupt cis-regulatory landscapes in vertebrates.
abstract: In multicellular organisms, cis-regulation controls gene expression in space and time. Despite the essential implication of cis-regulation in the development and evolution of organisms and in human diseases, our knowledge about regulatory sequences largely derives from analyzing their activity individually and outside their genomic context. Indeed, the contribution of these sequences to the expression of their target genes in their genomic context is still largely unknown. Here we present a novel genetic screen designed to visualize and interrupt gene regulatory landscapes in vertebrates. In this screen, based on the random insertion of an engineered Tol2 transposon carrying a strong insulator separating two fluorescent reporter genes, we isolated hundreds of zebrafish lines containing insertions that disrupt the cis-regulation of tissue-specific expressed genes. We therefore provide a new easy-to-handle tool that will help to disrupt and chart the regulatory activity spread through the vast noncoding regions of the vertebrate genome.
authors.affiliation: Centro Andaluz de Biología del Desarrollo (CABD), CSIC-Universidad Pablo de Olavide-Junta de Andalucía, Seville 41013, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Bessa
authors.forename: Mario
authors.initials: M
authors.lastname: Luengo
authors.forename: Solangel
authors.initials: S
authors.lastname: Rivero-Gil
authors.forename: Ana
authors.initials: A
authors.lastname: Ariza-Cosano
authors.forename: António H F
authors.initials: AH
authors.lastname: Maia
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Ruiz-Ruano
authors.forename: Pablo
authors.initials: P
authors.lastname: Caballero
authors.forename: Silvia
authors.initials: S
authors.lastname: Naranjo
authors.forename: Jaime J
authors.initials: JJ
authors.lastname: Carvajal
authors.forename: José Luis
authors.initials: JL
authors.lastname: Gómez-Skarmeta
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004251:DNA Transposable Elements / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005453:Fluorescence; D017930:Genes, Reporter / Q000502:physiology; D016678:Genome; D006801:Humans; D038101:Insulator Elements; D051379:Mice; D016254:Mutagenesis, Insertional / Q000379:methods*; D014714:Vertebrates / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements