Heat-inducible vectors for use in gene therapy.
The objectives of this study were to quantity and compare the activities of a minimal heat shock (HS) promoter and other promoters used in gene therapy applications, and to identify strategies to amplify the heat inducibility of therapeutic genes. Human tumour cells were transiently or stably transfected with the HS promoter driving expression of reporter genes. HS promoter activity was induced transiently, with maximum activity 16-24 h after HS, and was dependent on temperature. The activity of the minimal HS promoter was similar, after 42 degrees C HS for 1 h, to that of the cytomegalovirus (CMV) promoter. To determine if the HS promoter could be used to activate a second conditional promoter, cells were transiently transfected with vectors containing both the HS and human immunodeficiency virus type 1 (HIV1) promoters. When the IL-2 gene was placed downstream of the HIV1 promoter. IL-2 production was temperature-independent. The addition of the HIV tat gene downstream of the HS promoter caused IL-2 to be induced more than 3 fold after a single 42 degrees C HS. These data indicate that the minimal HS promoter, following activation by clinically attainable temperatures (< or = 42 degrees C), can drive expression of therapeutic genes at levels comparable to the CMV promoter and be used in conjunction with a second conditional promoter to drive temperature-dependent, gene expression.
Gerner
E W
EW
Department of Radiation Oncology, Cancer Biology Section, Arizona Cancer Center, The University of Arizona, Tucson 85724, USA. egerner@azcc.arizona.edu
Hersh
E M
EM
Pennington
M
M
Tsang
T C
TC
Harris
D
D
Vasanwala
F
F
Brailey
J
J
D015756:Genes, tat; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D006979:Hyperthermia, Induced; D007376:Interleukin-2 / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D014407:Tumor Cells, Cultured
D018840:HSP70 Heat-Shock Proteins; D007376:Interleukin-2; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins