PUBLICATION

Efficient transposition of the Tol2 transposable element from a single-copy donor in zebrafish

Authors
Urasaki, A., Asakawa, K., and Kawakami, K.
ID
ZDB-PUB-081217-17
Date
2008
Source
Proceedings of the National Academy of Sciences of the United States of America   105(50): 19827-19832 (Journal)
Registered Authors
Kawakami, Koichi
Keywords
heat shock, insertional mutagenesis, local hopping, nup214, transposon
MeSH Terms
  • Microinjections
  • Promoter Regions, Genetic
  • Transposases/genetics
  • DNA Transposable Elements/genetics*
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/metabolism
  • Male
  • Female
  • Mutagenesis, Insertional/methods*
  • Base Sequence
  • Molecular Sequence Data
  • Germ Cells
  • HSP70 Heat-Shock Proteins/genetics
  • Animals, Genetically Modified
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Animals
(all 17)
PubMed
19060204 Full text @ Proc. Natl. Acad. Sci. USA
Abstract
The Tol2 transposable element is a powerful genetic tool in model vertebrates and has been used for transgenesis, insertional mutagenesis, gene trapping, and enhancer trapping. However, an in vivo transposition system using Tol2 has not yet been developed. Here we report the in vivo Tol2 transposition system in a model vertebrate, zebrafish. First, we constructed transgenic zebrafish that carried single-copy integrations of Tol2 on the genome and injected transposase mRNA into one-cell stage embryos. The Tol2 insertions were mobilized efficiently in the germ lineage. We then mobilized an insertion of the Tol2 gene trap construct in the nup214 gene, which caused a recessive lethal mutant phenotype, and demonstrated that this method is applicable to the isolation of revertants from a transposon insertional mutant. Second, we constructed transgenic fish carrying the transposase cDNA under the control of the hsp70 promoter. Double-transgenic fish containing the transposase gene and a single-copy Tol2 insertion were treated with heat shock at the adult stage. We found that transposition can be induced efficiently in the male germ cells. We analyzed new integration sites and found that the majority (83%) of them were mapped on chromosomes other than the transposon donor chromosomes and that 9% of local hopping events mapped less than 300 kb away from the donor loci. Our present study demonstrates that the in vivo Tol2 transposition system is useful for creating genome-wide insertions from a single-copy donor and should facilitate functional genomics and transposon biology in vertebrates.
Genes / Markers
Figures
No images available
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
nkgsag37bGt/nkgsag37bGtstandard conditionsDay 5
1 - 1 of 1
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
nkfuji28TgTransgenic Insertion
    nkfuji70TgTransgenic Insertion
      nkgsag37bGtTransgenic Insertion
        nkhg6d-mh-1-3EtTransgenic Insertion
          nkhg6d-mh-1-4EtTransgenic Insertion
            nkhg6d-mh-1-5EtTransgenic Insertion
              nkhg6d-mh-1-6aEtTransgenic Insertion
                nkhg6d-mh-1-6bEtTransgenic Insertion
                  nkhg6d-mh-1-7EtTransgenic Insertion
                    nkhg6d-mh-1-8EtTransgenic Insertion
                      1 - 10 of 111
                      Show
                      Human Disease / Model
                      No data available
                      Sequence Targeting Reagents
                      Fish
                      Antibodies
                      Orthology
                      Engineered Foreign Genes
                      Marker Marker Type Name
                      DsRedEFGDsRed
                      EGFPEFGEGFP
                      1 - 2 of 2
                      Show
                      Mapping
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                      Citation
                      Urasaki, A., Asakawa, K., and Kawakami, K. (2008) Efficient transposition of the Tol2 transposable element from a single-copy donor in zebrafish. Proceedings of the National Academy of Sciences of the United States of America. 105(50):19827-19832.