Visualizing gene expression in living mammals using a bioluminescent reporter.
Control of gene expression often involves an interwoven set of regulatory processes. As information regarding regulatory pathways may be lost in ex vivo analyses, we used bioluminescence to monitor gene expression in living mammals. Viral promoters fused to firefly luciferase as transgenes in mice allowed external monitoring of gene expression both superficially and in deep tissues. In vivo bioluminescence was detectable using either intensified or cooled charge-coupled device cameras, and could be detected following both topical and systemic delivery of substrate. In vivo control of the promoter from the human immunodeficiency virus was demonstrated. As a model for DNA-based therapies and vaccines, in vivo transfection of a luciferase expression vector (SV-40 promoter and enhancer controlling expression) was detected. We conclude that gene regulation, DNA delivery and expression can now be noninvasively monitored in living mammals using a luciferase reporter. Thus, real-time, noninvasive study of gene expression in living animal models for human development and disease is possible.
Contag
C H
CH
Department of Pediatrics, Stanford University School of Medicine, CA 94305-5119, USA. ccontag@cmgm.stanford.edu
Spilman
S D
SD
Contag
P R
PR
Oshiro
M
M
Eames
B
B
Dennery
P
P
Stevenson
D K
DK
Benaron
D A
DA
D000818:Animals; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D016325:HIV Long Terminal Repeat; D006801:Humans; D019169:Jurkat Cells; D008156:Luciferases / Q000235:genetics; D049449:Luminescence; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic
D008156:Luciferases