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Closed Access item Zebrafish Enhancer Detection (ZED) Vector: A New Tool to Facilitate Transgenesis and the Functional Analysis of cis-Regulatory Regions in Zebrafish

Authors:Bessa, José
Tena, Juan J.
Calle-Mustienes, Elisa de la
Fernández-Miñán, Ana
Naranjo, Silvia
Fernández, Almudena
Montoliu, Lluís
Akalin, Altuna
Lenhard, Boris
Casares, Fernando
Gómez-Skarmeta, José Luis
Keywords:Zebrafish, Insulators, Transgenesis
Issue Date:Sep-2009
Publisher:John Wiley & Sons
Citation:Developmental Dynamics
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 cisregulatory 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.
Publisher version (URL):http://dx.doi.org/ 10.1002/dvdy.22051
Appears in Collections:(CNB) Artículos

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