Inducible gene expression by retrovirus-mediated transfer of a modified tetracycline-regulated system.
The ability to regulate gene expression via exogenous stimuli will facilitate the study of gene functions in mammalian cells. In the present study, we modified the tetracycline-controlled inducible system by the addition of the ligand-binding domain of the estrogen receptor to the carboxy terminus of the tTA transactivator. A single retroviral vector can transduce both the transactivator gene and the gene of interest controlled by the tTA-inducible promoter into mammalian cells. We show that cell lines expressing the transactivator can readily be established and that expression of the gene of interest depends on the removal of tetracycline and the addition of estrogen. By using this system, cell lines with inducible expression of the G protein of vesicular stomatitis virus, a potentially toxic gene product, were established. The combination of a powerful inducible system and retrovirus-mediated gene transfer can not only be used to study gene function but may also be applied in the future to clinical trials in human gene therapy.
Iida
A
A
Department of Pediatrics, City of Hope National Medical Center, Duarte, California 91010-3000, USA.
Chen
S T
ST
Friedmann
T
T
Yee
J K
JK
D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D004967:Estrogens / Q000494:pharmacology; D005354:Fibrosarcoma; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney; D008562:Membrane Glycoproteins; D008979:Moloney murine leukemia virus / Q000235:genetics; D011401:Promoter Regions, Genetic; D011960:Receptors, Estrogen / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012190:Retroviridae; D013752:Tetracycline / Q000494:pharmacology*; D013753:Tetracycline Resistance / Q000235:genetics*; D015534:Trans-Activators / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000096:biosynthesis*
D004967:Estrogens; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D015500:Chloramphenicol O-Acetyltransferase; D013752:Tetracycline