PUBLICATION
Gene-Gun-Mediated Transfer of Reporter Genes to Somatic Zebrafish (Danio rerio) Tissues
- Authors
- Torgersen, J., Collas, P., and Aleström, P.
- ID
- ZDB-PUB-100317-1
- Date
- 2000
- Source
- Marine biotechnology (New York, N.Y.) 2(3): 293-300 (Journal)
- Registered Authors
- Aleström, Peter, Collas, Philippe, Torgersen, Jacob
- Keywords
- gene gun, DNA transfer, expression, zebrafish, luciferase, GFP
- MeSH Terms
- none
- PubMed
- 10852809 Full text @ Mar. Biotechnol.
Citation
Torgersen, J., Collas, P., and Aleström, P. (2000) Gene-Gun-Mediated Transfer of Reporter Genes to Somatic Zebrafish (Danio rerio) Tissues. Marine biotechnology (New York, N.Y.). 2(3):293-300.
Abstract
We describe the use of gene-gun-mediated transfer of luciferase and green fluorescent protein (GFP) reporter genes in zebrafish (Danio rerio). Optimization of DNA transfer parameters indicated highest overall luciferase expression in epidermis and dermis using 1-µm microcarriers and 1 µg of pCMVL plasmid DNA at a delivery pressure of 200 psi. Time course studies revealed luciferase activity peaking at 18 hours and decreasing to 30% of the maximum at day 8 after DNA transfer. Onset of reporter gene (GFP) expression was detected at 13 minutes after DNA delivery, and by 65 minutes approximately 100% of the cells in the target area exhibited GFP expression. No germline association or integration events were detected in a screen of approximately 250,000 zebrafish sperm cells by fluorescence in situ hybridization at 15 or 30 days after delivery of 1 µg of pCMVL DNA, suggesting incidental male germline integration should not be considered as a risk factor when using the biolistic DNA delivery parameters and target tissues described.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping