Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system

Significance Bioluminescence is generated by luciferases that oxidize a specific luciferin. The enzymes involved in the synthesis of the luciferin from widespread cellular metabolites have so far been identified for only 2 bioluminescence systems, those of bacteria and fungi. In these cases, the complete reaction cascade is genetically encodable, meaning that heterologous expression of the corresponding genes can potentially produce autonomous bioluminescence in cell types other than the bacterial or fungal host cells. However, the light levels achieved in mammalian cells so far are not sufficient for single-cell applications. Here we present autonomous bioluminescence images of single mammalian cells by coexpression of the genes encoding the 6 enzymes from the bacterial bioluminescence system.