PUBLICATION
Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon
- Authors
- Davidson, A.E., Balciunas, D., Mohn, D., Shaffer, J., Hermanson, S., Sivasubbu, S., Cliff, M.P., Hackett, P.B., and Ekker, S.C.
- ID
- ZDB-PUB-031111-13
- Date
- 2003
- Source
- Developmental Biology 263(2): 191-202 (Journal)
- Registered Authors
- Balciunas, Darius, Ekker, Stephen C., Hackett, Perry B., Hermanson, Spencer, Mohn, Deanna, Sivasubbu, Sridhar
- Keywords
- none
- MeSH Terms
-
- Animals
- DNA Transposable Elements*
- Gene Dosage
- Gene Expression
- Gene Transfer Techniques*
- Mutagenesis, Insertional/genetics*
- Organ Specificity
- Promoter Regions, Genetic
- Transposases/physiology
- Zebrafish/genetics*
- PubMed
- 14597195 Full text @ Dev. Biol.
Citation
Davidson, A.E., Balciunas, D., Mohn, D., Shaffer, J., Hermanson, S., Sivasubbu, S., Cliff, M.P., Hackett, P.B., and Ekker, S.C. (2003) Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon. Developmental Biology. 263(2):191-202.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping