Optical imaging of Renilla luciferase, synthetic Renilla luciferase, and firefly luciferase reporter gene expression in living mice.
We have recently demonstrated that Renilla luciferase (Rluc) is a promising bioluminescence reporter gene that can be used for noninvasive optical imaging of reporter gene expression in living mice, with the aid of a cooled charged couple device (CCD) camera. In the current study, we explore the expression of a novel synthetic Renilla luciferase reporter gene (hRluc) in living mice, which has previously been reported to be a more sensitive reporter than native Rluc in mammalian cells. We explore the strategies of simultaneous imaging of both Renilla luciferase enzyme (RL) and synthetic Renilla luciferase enzyme (hRL):coelenterazine (substrate for RL/hRL) in the same living mouse. We also demonstrate that hRL:coelenterazine can yield a higher signal when compared to Firefly luciferase enzyme (FL): D-Luciferin, both in cell culture studies and when imaged from cells at the surface and from lungs of living mice. These studies demonstrate that hRluc should be a useful primary reporter gene with high sensitivity when used alone or in conjunction with other bioluminescence reporter genes for imaging in living rodents.
Bhaumik
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The Crump Institute for Molecular Imaging, 700 Westwood Boulevard, Los Angeles, California 90095-1700, USA.
Lewis
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XZ
Gambhir
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D000818:Animals; D037421:Anthozoa / Q000201:enzymology*; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D001517:Coleoptera / Q000201:enzymology*; D049454:Firefly Luciferin / Q000493:pharmacokinetics*; D017930:Genes, Reporter / Q000235:genetics; D005910:Glioma / Q000378:metabolism; D006801:Humans; D007093:Imidazoles; D007527:Isoenzymes; D007668:Kidney / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008657:Metabolic Clearance Rate; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D009447:Neuroblastoma / Q000378:metabolism; D009928:Organ Specificity; D011719:Pyrazines / Q000493:pharmacokinetics*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D014018:Tissue Distribution
D007093:Imidazoles; D007527:Isoenzymes; D011719:Pyrazines; C017144:coelenterazine; D049454:Firefly Luciferin; D008156:Luciferases