Abstract
In vivo bioluminescence imaging (BLI), which is based on luminescence emitted by the luciferase–luciferin reaction, has enabled continuous monitoring of various biochemical processes in living animals. Bright luminescence with a high signal-to-background ratio, ideally red or near-infrared light as the emission maximum, is necessary for in vivo animal experiments. Various attempts have been undertaken to achieve this goal, including genetic engineering of luciferase, chemical modulation of luciferin, and utilization of bioluminescence resonance energy transfer (BRET). In this review, we overview a recent advance in the development of a bioluminescence system for in vivo BLI. We also specifically examine the improvement in bioluminescence intensity by mutagenic or chemical modulation on several beetle and marine luciferase bioluminescence systems. We further describe that intramolecular BRET enhances luminescence emission, with recent attempts for the development of red-shifted bioluminescence system, showing great potency in in vivo BLI. Perspectives for future improvement of bioluminescence systems are discussed.
Highlights
Quantitative monitoring of individual biomolecular reactions and cellular behaviors is necessary to elucidate the dynamic and complicated functions of biological phenomena
bioluminescence imaging (BLI) is based on the catalytic activity of luciferase enzymes, which oxidize the substrate called luciferin to generate an excited-state molecule that emits bioluminescence
BLI enables highly sensitive imaging in living organisms because it requires no external light for light emission
Summary
Quantitative monitoring of individual biomolecular reactions and cellular behaviors is necessary to elucidate the dynamic and complicated functions of biological phenomena. In vivo bioluminescence imaging (BLI) has enabled visualization of biological processes in intact living organisms, providing abundant quantitative, spatiotemporal information that is far beyond the reach of conventional in vitro assays. BLI is based on the catalytic activity of luciferase enzymes, which oxidize the substrate called luciferin to generate an excited-state molecule that emits bioluminescence. All luciferins require chemical reactions to emit bioluminescence: the oxidation of luciferins by luciferases to yield excited-state oxyluciferin, and relaxation to the ground state with photon emission. Despite their similarity, the color and intensity of the emitted bioluminescence and the dependency on pH and other molecules (e.g., ATP) differ among luciferase–luciferin pairs. BRET ratio: 34.0 BRET ratio: 4.0 Via SNAP-tag or HaloTag Via SNAP-tag or HaloTag Via SNAP-tag or HaloTag Via SNAP-tag or HaloTag BRET ratio: 0.70 BRET ratio: 1.2 BRET ratio: 2.3 BRET ratio: 1.32 BRET ratio: 13.3
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