Gene-Gun-Mediated Transfer of Reporter Genes to Somatic Zebrafish (Danio rerio) Tissues |
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Authors: | Jacob Torgersen Philippe Collas Peter Aleström |
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Institution: | (1) Department of Biochemistry, Physiology and Nutrition, Norwegian School of Veterinary Science, P.O. Box 8146 Dep., N-0033 Oslo, Norway, NO;(2) Institute of Medical Biochemistry, Faculty of Medicine, University of Oslo, Norway, NO |
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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.
Received August 20, 1999; accepted January 6, 2000. |
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Keywords: | : gene gun DNA transfer expression zebrafish luciferase GFP |
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