title: Zebrafish enhancer detection (ZED) vector: a new tool to facilitate transgenesis and the functional analysis of cis-regulatory regions in zebrafish.
abstract: The identification and characterization of the regulatory activity of genomic sequences is crucial for understanding how the information contained in genomes is translated into cellular function. The cis-regulatory sequences control when, where, and how much genes are transcribed and can activate (enhancers) or repress (silencers) gene expression. Here, we describe a novel Tol2 transposon-based vector for assessing enhancer activity in the zebrafish (Danio rerio). This Zebrafish Enhancer Detector (ZED) vector harbors several key improvements, among them a sensitive and specific minimal promoter chosen for optimal enhancer activity detection, insulator sequences to shield the minimal promoter from position effects, and a positive control for transgenesis. Additionally, we demonstrate that highly conserved noncoding sequences homologous between humans and zebrafish largely with enhancer activity largely retain their tissue-specific enhancer activity during vertebrate evolution. More strikingly, insulator sequences from mouse and chicken, but not conserved in zebrafish, maintain their insulator capacity when tested in this model.
authors: Centro Andaluz de Biología del Desarrollo, CSIC-Universidad Pablo de Olavide, Campus UPO, Seville, Spain. jcribbes@upo.es
authors: José
authors: J
authors: Bessa
authors: Juan J
authors: JJ
authors: Tena
authors: Elisa
authors: E
authors: de la Calle-Mustienes
authors: Ana
authors: A
authors: Fernández-Miñán
authors: Silvia
authors: S
authors: Naranjo
authors: Almudena
authors: A
authors: Fernández
authors: Lluis
authors: L
authors: Montoliu
authors: Altuna
authors: A
authors: Akalin
authors: Boris
authors: B
authors: Lenhard
authors: Fernando
authors: F
authors: Casares
authors: José Luis
authors: JL
authors: Gómez-Skarmeta
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D051379:Mice; D015027:Zebrafish / Q000235:genetics*