Development of a VSV-G protein pseudotyped retroviral vector system expressing dominant oncogenes from a lacO-modified inducible LTR promoter

Gene. 1996 Dec 5;182(1-2):145-50. doi: 10.1016/s0378-1119(96)00536-7.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, Polyomavirus Transforming / metabolism
  • Blotting, Northern
  • Chloramphenicol O-Acetyltransferase / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblasts
  • Gene Expression Regulation / genetics
  • Genes, Reporter / genetics
  • Genes, Viral
  • Genes, ras / genetics
  • Genetic Vectors / genetics*
  • Gentamicins / pharmacology
  • Humans
  • Isopropyl Thiogalactoside / pharmacology
  • Oncogenes / genetics
  • Plasmids / genetics
  • Retroviridae / genetics*
  • Transformation, Genetic / genetics

Substances

  • Antigens, Polyomavirus Transforming
  • Gentamicins
  • Isopropyl Thiogalactoside
  • antibiotic G 418
  • Chloramphenicol O-Acetyltransferase