Generation of packaging cell lines for pseudotyped retroviral vectors of the G protein of vesicular stomatitis virus by using a modified tetracycline inducible system.
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.
Chen
S T
ST
Department of Pediatrics, University of California at San Diego, La Jolla 92093, USA.
Iida
A
A
Guo
L
L
Friedmann
T
T
Yee
J K
JK
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D004958:Estradiol / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D006801:Humans; D007700:Kinetics; D008156:Luciferases / Q000096:biosynthesis; D008562:Membrane Glycoproteins; D051379:Mice; D051381:Rats; D011960:Receptors, Estrogen / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012190:Retroviridae; D013752:Tetracycline / Q000494:pharmacology*; D013937:Thymidine Kinase / Q000096:biosynthesis; D015534:Trans-Activators / Q000096:biosynthesis*; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000096:biosynthesis*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D004958:Estradiol; D008156:Luciferases; D013937:Thymidine Kinase; D013752:Tetracycline