Development and Applications of Bioluminescent and Chemiluminescent Reporters and Biosensors

Abstract

Although fluorescent reporters and biosensors have become indispensable tools in biological and biomedical fields, fluorescence measurements require external excitation light, thereby limiting their use in thick tissues and live animals. Bioluminescent reporters and biosensors may potentially overcome this hurdle because they use enzyme-catalyzed exothermic biochemical reactions to generate excited-state emitters. This review first introduces the development of bioluminescent reporters, and next, their applications in sensing biological changes in vitro and in vivo as biosensors. Lastly, we discuss chemiluminescent sensors that produce photons in the absence of luciferases. This review aims to explore fundamentals and experimental insights and to emphasize the yet-to-be-reached potential of next-generation luminescent reporters and biosensors.

Examining multiple cellular pathways at once using multiplex hextuple luciferase assaying

“…To reduce variability of measurements as much as possible, stable cell lines could be generated by the targeted integration of all multiplex reporters at the same defined site in the genome, such as a safe harbor site 50 , resulting in the neutralization of genomic position effects. Cells with such genome-integrated luciferase reporters can then be further used to explore luciferase multiplexing towards in vivo bioluminescence applications [51][52][53] . Another concern may be potential cross-talk between the reporters integrated into the same plasmid by synthetic biology.…”

“…In future work, more luciferases could be integrated to expand the capabilities of this assay. Approaches to accomplish this include molecular modification of existing luciferases into regions of emission spectra not yet covered by the D-Luciferin-or coelenterazine-responsive luciferases or investigation of luciferases that utilize other substrates (such as vargulin and furimazine) 13,51 . Additional luciferases can be easily incorporated in our multiplex pipeline by synthetic assembly [25][26][27] .…”

Recent Progress in Optical Sensors for Biomedical Diagnostics

“…Bioluminescence is the production and emission of light by living organisms for defence, prey or communication [181]. The biochemical reaction involved in the production of light is the oxidation of a luciferin molecule (lucifer means "light-bearer") by a luciferase enzyme [182]. Luciferins can be small molecules like coelenterazine, furimazine, D-luciferin (Figure 9a) with few cyclic structures as well as larger photoproteins such as aequorin, symplectin, pholasin, berovin, obelin and clytin (Figure 9b).…”

Luciferase‐Induced Photouncaging: Bioluminolysis

“…Bioluminescent enzymes,c ollectively known as luciferases, have been broadly used as reporters in biological applications ranging from cell-based assays to in vivo imaging. [1][2][3][4] Luciferases catalyze the oxidation of chemical substrates to generate photons,with or without recourse to cofactors. [5] Compared to conventional fluorescence imaging, bioluminescence imaging is ultrasensitive,s hows high signal-to-noise ratios,a nd does not cause phototoxicity.B ioluminescence resonance energy transfer (BRET) thus attracted significant attention in the study of dynamic processes in biological systems such as protein-protein interactions,signaling events,and metabolite or drug sensors.…”