Zebrafish enhancer detection (ZED) vector: A new tool to facilitate transgenesis and the functional analysis of cis-regulatory regions in zebrafish
Article first published online: 3 AUG 2009
Copyright © 2009 Wiley-Liss, Inc.
Special Issue: Special Focus on Visual Systems
Volume 238, Issue 9, pages 2409–2417, September 2009
How to Cite
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. (2009), Zebrafish enhancer detection (ZED) vector: A new tool to facilitate transgenesis and the functional analysis of cis-regulatory regions in zebrafish. Dev. Dyn., 238: 2409–2417. doi: 10.1002/dvdy.22051
- Issue published online: 13 AUG 2009
- Article first published online: 3 AUG 2009
- Manuscript Accepted: 30 JUN 2009
- Spanish Ministry of Education and Science. Grant Numbers: BFU2007-60042/BMC, Petri PET2007_0158, CSD2007-00008
- Junta de Andalucía. Grant Numbers: Proyecto de Excelencia CVI260, CVI-3488
- CSIC, Universidad Pablo de Olavide and Junta de Andalucía
- enhancer detection;
- gene regulation;
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. Developmental Dynamics 238:2409–2417, 2009. © 2009 Wiley-Liss, Inc.