Use of an autonomous parvovirus vector for selective transfer of a foreign gene into transformed human cells of different tissue origins and its expression therein.
In this work, we report the transduction of a chloramphenicol acetyltransferase (CAT) reporter gene into a variety of normal and transformed human cells of various tissue origins. The vector used was MVM/P38cat, a recombinant of the prototype strain of the autonomous parvovirus minute virus of mice (MVMp). The CAT gene was inserted into the capsid-encoding region of the infectious molecular clone of MVMp genome, under the control of the MVM P38 promoter. When used to transfect permissive cells, the MVM/P38cat DNA was efficiently replicated and expressed the foreign CAT gene at high levels. By cotransfecting with a helper plasmid expressing the capsid proteins, it was possible to produce mixed virus stocks containing MVM/P38cat infectious particles and variable amounts of recombinant MVM. MVM/P38cat viral particles were successfully used to transfer the CAT gene and to express it in a variety of human cells. Both viral DNA replication and P38-driven CAT expression were achieved in fibroblasts, epithelial cells, T lymphocytes, and macrophages in a transformation-dependent way, but with an efficiency depending on the cell type. In transformed B lymphocytes, however, the vector was not replicated, nor did it express the CAT gene.
Dupont
F
F
Department of Molecular Biology, Université Libre de Bruxelles, Rhode Saint Genèse, Belgium.
Tenenbaum
L
L
Guo
L P
LP
Spegelaere
P
P
Zeicher
M
M
Rommelaere
J
J
D000818:Animals; D002461:Cell Line, Transformed; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D004261:DNA Replication; D004847:Epithelial Cells; D004848:Epithelium / Q000382:microbiology; D005347:Fibroblasts / Q000166:cytology / Q000382:microbiology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000382:microbiology; D008915:Minute Virus of Mice / Q000235:genetics* / Q000254:growth & development; D013601:T-Lymphocytes / Q000166:cytology / Q000382:microbiology; D014161:Transduction, Genetic; D014162:Transfection; D014779:Virus Replication
D015500:Chloramphenicol O-Acetyltransferase