Reporter gene expression in fish following cutaneous infection with pantropic retroviral vectors

A central issue in gene delivery systems is choosing promoters that will direct defined and sustainable levels of gene expression. Pantropic retroviral vectors provide a means to insert genes into either somatic or germline cells. In this study, we focused on somatic cell infection by evaluating the...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2001-06, Vol.3 (Supplement 1), p.S81-S087
Main Authors: Paul, T A, Burns, J C, Shike, H, Getchell, R, Bowser, P R, Whitlock, K E, Casey, J W
Format: Article
Language:English
Subjects:
Biotechnology
Cell lines
Cell proliferation
Cells
Cytomegalovirus
Danio rerio
Deoxyribonucleic acid
DNA
Expression vectors
Fish
Fish skin
Fishing lines
Freshwater
Freshwater fishes
Gene expression
Gene transfer
Genomes
Lepomis macrochirus
Leukemia
Long terminal repeat
Marine fishes
Neoplasms
Proliferation
Promoters
Reporter gene
Sander vitreus
Sarcoma
Tissue
transgenic fish
Vectors
Viruses
Zebrafish
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Summary:A central issue in gene delivery systems is choosing promoters that will direct defined and sustainable levels of gene expression. Pantropic retroviral vectors provide a means to insert genes into either somatic or germline cells. In this study, we focused on somatic cell infection by evaluating the activity of 3 promoters inserted by vectors into fish cell lines and fish skin using pantropic retroviruses. In bluegill and zebrafish cell lines, the highest levels of luciferase expression were observed from the 5' murine leukemia virus long terminal repeat of the retroviral vector. The Rous sarcoma virus long terminal repeat and cytomegalovirus early promoter, as internal promoters, generated lower levels of luciferase. Luciferase reporter vectors infected zebrafish skin, as measured by the presence of viral DNA, and expressed luciferase. We infected developing walleye dermal sarcomas with retroviral vectors to provide an environment with enhanced cell proliferation, a condition necessary for integration of the provirus into the host genome. We demonstrated a 4-fold to 7-fold increase in luciferase gene expression in tumor tissue over infections in normal walleye skin.
ISSN:1436-2228
1436-2236
DOI:10.1007/s10126-001-0029-y