Somatic Mutagenesis with a Sleeping Beauty Transposon System Leads to Solid Tumor Formation in Zebrafish

Thumbnail Image
Date
2011-04-01
Authors
Hatler, Julia
Kuang, Zianyan
Liao, Hsin-Kai
Nannapaneni, Kishore
Noack Watt, Kristin
Uhl, Juli
Largaespada, David
Vollbrecht, Erik
Major Professor
Advisor
Committee Member
Journal Title
Journal ISSN
Volume Title
Publisher
Authors
Person
McGrail, Maura
Associate Professor
Person
Essner, Jeffrey
Professor
Research Projects
Organizational Units
Organizational Unit
Genetics, Development and Cell Biology

The Department of Genetics, Development, and Cell Biology seeks to teach subcellular and cellular processes, genome dynamics, cell structure and function, and molecular mechanisms of development, in so doing offering a Major in Biology and a Major in Genetics.

History
The Department of Genetics, Development, and Cell Biology was founded in 2005.

Related Units

Journal Issue
Is Version Of
Versions
Series
Abstract

Large-scale sequencing of human cancer genomes and mouse transposon-induced tumors has identified a vast number of genes mutated in different cancers. One of the outstanding challenges in this field is to determine which genes, when mutated, contribute to cellular transformation and tumor progression. To identify new and conserved genes that drive tumorigenesis we have developed a novel cancer model in a distantly related vertebrate species, the zebrafish, Danio rerio. The Sleeping Beauty (SB) T2/Onc transposon system was adapted for somatic mutagenesis in zebrafish. The carp ß-actin promoter was cloned into T2/Onc to create T2/OncZ. Two transgenic zebrafish lines that contain large concatemers of T2/ OncZ were isolated by injection of linear DNA into the zebrafish embryo. The T2/OncZ transposons were mobilized throughout the zebrafish genome from the transgene array by injecting SB11 transposase RNA at the 1-cell stage. Alternatively, the T2/OncZ zebrafish were crossed to a transgenic line that constitutively expresses SB11 transposase. T2/ OncZ transposon integration sites were cloned by ligation-mediated PCR and sequenced on a Genome Analyzer II. Between 700–6800 unique integration events in individual fish were mapped to the zebrafish genome. The data show that introduction of transposase by transgene expression or RNA injection results in an even distribution of transposon reintegration events across the zebrafish genome. SB11 mRNA injection resulted in neoplasms in 10% of adult fish at ,10 months of age. T2/OncZ-induced zebrafish tumors contain many mutated genes in common with human and mouse cancer genes. These analyses validate our mutagenesis approach and provide additional support for the involvement of these genes in human cancers. The zebrafish T2/OncZ cancer model will be useful for identifying novel and conserved genetic drivers of human cancers.

Comments

This article is from PLoS One 6 (2011): e18826, doi:10.1371/journal.pone.0018826. Posted with permission.

Description
Keywords
Citation
DOI
Copyright
Sat Jan 01 00:00:00 UTC 2011
Collections