Engineering Autobioluminescent Eukaryotic Cells as Tools for Environmental and Biomedical Surveillance

Cells engineered to express bioluminescence generate photons through a biochemical reaction catalyzed by a luciferase enzyme and a luciferin substrate. Conventional bioluminescent imaging approaches require that the luciferin substrate be added to the cell externally since it cannot be generated intracellularly. Thus, these cells remain “dark, " only emitting their bioluminescent signal in concert with the extracellular addition of the chemical substrate. Autobioluminescence represents an alternative signaling strategy whereby a synthetic bacterial luciferase, optimized for expression in eukaryotic cells, is used to sustain continuous photon emission without the necessary external addition of a substrate to drive reaction kinetics. This substrate-independent generation of light thereby allows autobioluminescent cells to be imaged at any longitudinal time scale desired to generate real-time metabolic data under both highthroughput in vitro and small animal in vivo experimental set-ups for applications ranging from environmental monitoring to preclinical biomedical imaging.