A mobile insulator system to detect and disrupt cis-regulatory landscapes in vertebrates
- José Bessa1,3,
- Mario Luengo2,
- Solangel Rivero-Gil2,
- Ana Ariza-Cosano2,
- António H.F. Maia,
- Francisco J. Ruiz-Ruano,
- Pablo Caballero,
- Silvia Naranjo,
- Jaime J. Carvajal and
- José Luis Gómez-Skarmeta3
- Centro Andaluz de Biología del Desarrollo (CABD), CSIC-Universidad Pablo de Olavide–Junta de Andalucía, Seville 41013, Spain
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↵2 These authors contributed equally to this work.
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.
Footnotes
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↵3 Corresponding authors
E-mail Jose.Bessa{at}ibmc.up.pt
E-mail jlgomska{at}upo.es
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[Supplemental material is available for this article.]
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Article published online before print. Article, supplemental material, and publication date are at http://www.genome.org/cgi/doi/10.1101/gr.165654.113.
- Received August 26, 2013.
- Accepted November 20, 2013.
This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.
