Basic and Applied Aspects of Color Tuning of Bioluminescence Systems

Abstract

V. Viviani et al. [Biochemistry 38 (1999) 8271] were the first to succeed in cloning the red-emitting enzyme from the South American railroad worm, which is the only bioluminescent organism known to emit a red-colored light. The application of red bioluminescence has been our goal because the transmittance of longer-wavelength light is superior to that of the other colors for visualization of biological functions in living cells. Now, different color luciferases, which emit with wavelength maxima ranging from 400 to 630 nm, are available and are being used. For example, based on different color luciferases, Nakajima et al. developed a tricolor reporter in vitro assay system based on these different color luciferases in which the expression of three genes can be monitored simultaneously. On the other hand, bioluminescence resonance energy transfer (BRET) is a natural phenomenon caused by the intermolecular interaction between a bioluminescent protein and a fluorophore on a second protein, resulting in the light from the bioluminescence reaction having the spectrum of the fluorophore. Otsuji et al. [Anal. Biochem. 329 (2004) 230] showed that the change in the efficiency of energy transfer in intramolecular BRET can quantify cellular functions in living cells. In this review, I introduce the basic mechanisms of color tuning in bioluminescent systems and new applications based on color tuning in the life sciences.

Aqueous Near‐Infrared Fluorescent Composites Based on Apoferritin‐Encapsulated PbS Quantum Dots

“…To overcome this difficulty, the technique of BRET using a self-illuminating system has been developed. [384,385] Bioluminescence is an enzyme-catalysed reaction involving the enzyme luciferase, the most well known being firefly luciferase from the firefly Photinus pyralis. …”

Dual-Reporter Assay Using Two Secreted Luciferase Genes

“…Luciferase is used as a reporter enzyme to estimate gene expression in prokaryotic or eukaryotic cells because the amount of luciferase present is correlated with light intensity in the presence of excess luciferin (1). Furthermore, its sensitivity and range of linear responses are superior to those of other typical reporters (2).…”

Evolution of BRET Biosensors from Live Cell to Tissue-Scale In vivo Imaging

“…FLuc has been reported to be used in a BRET system in conjunction with red FPs like DsRed (25) as well as with non-protein fluorophores such as Cy3 and Cy3.5 (26). However, its bulky size of 61 kDa, an obligate dependence on Mg 2+ and ATP as its cofactors (27, 28) and finally, a low spectral resolution with the BRET partners reported so far, makes it a poor choice for BRET.…”