Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon

We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish ( Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expr...

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Bibliographic Details
Published in:Developmental biology 2003-11, Vol.263 (2), p.191-202
Main Authors: Davidson, Ann E, Balciunas, Darius, Mohn, Deanna, Shaffer, Jennifer, Hermanson, Spencer, Sivasubbu, Sridhar, Cliff, M.Pat, Hackett, Perry B, Ekker, Stephen C
Format: Article
Language:English
Subjects:
Animals
DNA Transposable Elements
Freshwater
Gene Dosage
Gene Expression
Gene Transfer Techniques
Insertional mutagenesis
Mutagenesis, Insertional - genetics
Organ Specificity
Promoter Regions, Genetic
Sleeping Beauty
Transgene expression
Transposases - physiology
Transposon
Zebrafish
Zebrafish - genetics
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Summary:We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish ( Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.
ISSN:0012-1606
1095-564X
DOI:10.1016/j.ydbio.2003.07.013