Novel gene switches for targeted and timed expression of proteins of interest.
This article reports on the construction and analysis in vitro and in vivo of novel gene switches that can be used to achieve spatial as well as temporal control over the expression of a transgene of interest. The switches are expected to be functional in virtually any tissue and cell type. They consist of (a) a foreign or modified transactivator expressed under the dual control of a promoter or promoter cassette that is responsive to heat and the transactivator and (b) a promoter responsive to the transactivator for controlling the transgene of interest. A preferred gene switch of this type incorporated a mifepristone-dependent transactivator. This gene switch could be activated by a transient heat treatment in the presence of mifepristone. Activity increased with the intensity of the activating heat treatment and was found to persist for more than 6 days. The gene switch was essentially inactive prior to an activating heat treatment, in the absence or presence of mifepristone. Activated gene switch could be silenced by removal/withdrawal of mifepristone.
Vilaboa
Nuria
N
Laboratorio de Metabolismo Oseo, Hospital Universitario La Paz, Paseo de la Castellana 261, 28046 Madrid, Spain.
Fenna
Mary
M
Munson
John
J
Roberts
Stephen M
SM
Voellmy
Richard
R
D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005812:Genes, Switch / Q000187:drug effects / Q000502:physiology*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000235:genetics*; D006358:Hot Temperature; D006801:Humans; D015735:Mifepristone / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*; D015534:Trans-Activators / Q000494:pharmacology*; D014158:Transcription, Genetic; D014162:Transfection
D006360:Heat-Shock Proteins; D015534:Trans-Activators; D015735:Mifepristone