Imaging cyclooxygenase-2 (Cox-2) gene expression in living animals with a luciferase knock-in reporter gene.
The cyclooxygenase-2 (Cox-2) gene plays a role in a variety of normal and pathophysiological conditions. Expression of the Cox-2 gene is induced in a broad range of cells, in response to many distinct stimuli. The ability to monitor and quantify Cox-2 expression noninvasively in vivo may facilitate a better understanding of the role of Cox-2, both in normal physiology and in different diseases. We generated a "knock-in" mouse in which the firefly luciferase reporter enzyme is expressed at the start site of translation of the endogenous Cox-2 gene. Correlation of luciferase and Cox-2 expression was confirmed in heterozygous Cox-2luc/+ mouse embryonic fibroblasts isolated from the knock-in mouse. In an acute sepsis model, following injection of interferon gamma and endotoxin, ex vivo imaging and Western blotting demonstrated coordinate Cox-2 and luciferase induction in multiple organs. Using both paw and air pouch inflammation models, we can monitor repeatedly localized luciferase expression in the same living mouse. Cox-2luc/+ knock-in mice should provide a valuable tool to analyze Cox-2 expression in many disease models.
Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, 341 Boyer Hall, 611 Charles E. Young Drive East, UCLA, Los Angeles, CA 90095, USA.
Tomo-O
TO
Ishikawa
Naveen K
NK
Jain
Makoto M
MM
Taketo
Harvey R
HR
Herschman
D000818:Animals; D052160:Benzothiazoles / Q000008:administration & dosage / Q000378:metabolism; D051546:Cyclooxygenase 2 / Q000096:biosynthesis* / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000201:enzymology; D048888:Fireflies / Q000235:genetics; D055879:Gene Knock-In Techniques / Q000379:methods; D017930:Genes, Reporter; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic; D015054:Zymosan / Q000008:administration & dosage
D052160:Benzothiazoles; C532924:D-luciferin; D015054:Zymosan; D008156:Luciferases; D051546:Cyclooxygenase 2