Generation of packaging cell lines for pseudotyped retroviral vectors of the G protein of vesicular stomatitis virus by using a modified tetracycline inducible system

Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10057-62. doi: 10.1073/pnas.93.19.10057.

Abstract

We have previously shown that the G protein of vesicular stomatitis virus (VSV-G) can be incorporated into the virions of retroviruses. Since expression of VSV-G is toxic to most mammalian cells, development of stable VSV-G packaging cell lines requires inducible VSV-G expression. We have modified the tetracycline-inducible system by fusing the ligand binding domain of the estrogen receptor to the carboxy terminus of a tetracycline-regulated transactivator. Using this system, we show that VSV-G expression is tetracycline-dependent and can be modulated by beta-estradiol. Stable packaging cell lines can readily be established and high-titer pseudotyped retroviral vectors can be generated upon induction of VSV-G expression.

Publication types

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

MeSH terms

  • 3T3 Cells
  • Animals
  • Cell Line
  • Estradiol / pharmacology*
  • Gene Expression / drug effects
  • Genetic Vectors*
  • Humans
  • Kinetics
  • Luciferases / biosynthesis
  • Membrane Glycoproteins*
  • Mice
  • Rats
  • Receptors, Estrogen / biosynthesis*
  • Recombinant Fusion Proteins / biosynthesis*
  • Retroviridae*
  • Tetracycline / pharmacology*
  • Thymidine Kinase / biosynthesis
  • Trans-Activators / biosynthesis*
  • Transcriptional Activation / drug effects
  • Transfection
  • Vesicular stomatitis Indiana virus / genetics*
  • Vesicular stomatitis Indiana virus / metabolism
  • Viral Envelope Proteins / biosynthesis*

Substances

  • G protein, vesicular stomatitis virus
  • Membrane Glycoproteins
  • Receptors, Estrogen
  • Recombinant Fusion Proteins
  • Trans-Activators
  • Viral Envelope Proteins
  • Estradiol
  • Luciferases
  • Thymidine Kinase
  • Tetracycline