Dual Reporter Bioluminescence Imaging with NanoLuc and Firefly Luciferase.

Bioluminescence imaging is a powerful, broadly utilized method for noninvasive imaging studies in cell-based assays and small animal models of normal physiology and multiple diseases. In combination with molecular engineering of cells and entire organisms using luciferase enzymes, bioluminescence imaging has enabled novel applications including studies of protein-protein interactions, ligand-receptor interactions, cell trafficking, and drug targeting in mouse models. We describe use of a novel luciferase enzyme derived from Oplophorus gracilirostris, NanoLuc, in cell-based assays bioluminescence imaging of tumor-bearing mice. We also combine NanoLuc with another luciferase enzyme, firefly luciferase, to image multiple signal transduction events in one imaging session.