ZFIN ID: ZDB-PUB-090807-14
Zebrafish enhancer detection (ZED) vector: A new tool to facilitate transgenesis and the functional analysis of cis-regulatory regions in zebrafish
Bessa, J., Tena, J.J., de la Calle-Mustienes, E., Fernández-Miñán, A., Naranjo, S., Fernández, A., Montoliu, L., Akalin, A., Lenhard, B., Casares, F., and Gómez-Skarmeta, J.L.
Date: 2009
Source: Developmental dynamics : an official publication of the American Association of Anatomists   238(9): 2409-2417 (Journal)
Registered Authors: Bessa, Jose, Casares, Fernando, de la Calle-Mustienes, Elisa, Gómez-Skarmeta, José Luis, Naranjo, Silvia, Tena, Juan
Keywords: enhancer detection, gene regulation, insulators, zebrafish, transgenesis
MeSH Terms:
  • Animals
  • Animals, Genetically Modified/genetics
  • Enhancer Elements, Genetic/genetics*
  • Gene Transfer Techniques*
  • Genetic Vectors/genetics*
  • Mice
  • Zebrafish/genetics*
PubMed: 19653328 Full text @ Dev. Dyn.
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.