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
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3T3 Cells
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Animals
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Cell Line
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Estradiol / pharmacology*
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Gene Expression / drug effects
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Genetic Vectors*
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Humans
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Kinetics
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Luciferases / biosynthesis
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Membrane Glycoproteins*
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Mice
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Rats
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Receptors, Estrogen / biosynthesis*
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Recombinant Fusion Proteins / biosynthesis*
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Retroviridae*
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Tetracycline / pharmacology*
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Thymidine Kinase / biosynthesis
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Trans-Activators / biosynthesis*
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Transcriptional Activation / drug effects
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Transfection
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Vesicular stomatitis Indiana virus / genetics*
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Vesicular stomatitis Indiana virus / metabolism
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Viral Envelope Proteins / biosynthesis*
Substances
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G protein, vesicular stomatitis virus
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Membrane Glycoproteins
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Receptors, Estrogen
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Recombinant Fusion Proteins
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Trans-Activators
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Viral Envelope Proteins
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Estradiol
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Luciferases
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Thymidine Kinase
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Tetracycline