Development of a VSV-G protein pseudotyped retroviral vector system expressing dominant oncogenes from a lacO-modified inducible LTR promoter.
We report the development of a retroviral vector system in which two dominant oncogenes are expressed inducibly in human cells using the lac repressor/lac operator regulatable promoter system. First, the parent vector, pLoCRNLo, was constructed to contain a retroviral long terminal repeat (LTR) promoter that has been modified by incorporation of a lac operator sequence (lacO). This promoter, LTRo, was shown to mediate IPTG-inducible cat expression in rat cells expressing the lac repressor. The pLoCRNLo backbone was used to develop the retroviral vector LoTPRRNLo which expresses SV40 T antigen and H-ras val12 oncogenes as a dicistronic unit separated by a poliovirus internal ribosome entry sequence (PO-IRES). LoPRRNLo retrovirus was produced as a VSV-G protein pseudotype and used to infect primary human cells, resulting in the efficient formation of transformed cell lines. Subsequent introduction into the transformed cells of the lac repressor, expressed from a second retroviral vector, MSCV-In(S), resulted in IPTG-responsive oncogene expression and cell growth. This vector system is useful for introducing multiple genes under inducible control into mammalian cells.
Wang
S
S
Department of Pediatrics, UCSD School of Medicine, La Jolla 92093-0634, USA.
Beattie
G M
GM
Hayek
A
A
Levine
F
F
D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D015152:Blotting, Northern; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts; D005786:Gene Expression Regulation / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D005814:Genes, Viral; D011905:Genes, ras / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D005839:Gentamicins / Q000494:pharmacology; D006801:Humans; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D009857:Oncogenes / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics
D000952:Antigens, Polyomavirus Transforming; D005839:Gentamicins; D007544:Isopropyl Thiogalactoside; C010680:antibiotic G 418; D015500:Chloramphenicol O-Acetyltransferase