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A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

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Luciferases with Tunable Emission Wavelengths.

Julien Hiblot^1,2, Qiuliyang Yu^1,2, Marina D B Sabbadini¹

¹Ecole Polytechnique Fédérale de Lausanne, EPFL, Institute of Chemical Sciences and Engineering, ISIC, NCCR in Chemical Biology, 1015, Lausanne, Switzerland.

Angewandte Chemie (International Ed. in English)

|September 24, 2017

Summary

Researchers created new luciferases with adjustable colors using chemical labeling. These engineered proteins, based on NanoLuc, can be modified to emit light at various wavelengths for diverse biological applications.

Keywords:

bioluminescent imaging luciferase self-labeling protein sensors therapeutic drug monitoring

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Area of Science:

Biochemistry
Molecular Biology
Biotechnology

Background:

Luciferases are enzymes that emit light.
Controlling luciferase emission color is crucial for multiplexed biological assays.
Existing luciferases have fixed emission spectra, limiting their application in complex systems.

Purpose of the Study:

To develop novel luciferases with tunable emission wavelengths.
To create versatile reporter systems for biological research.
To enable advanced bioimaging and biosensor development.

Main Methods:

Constructing chimeric luciferases by fusing NanoLuc with self-labeling tags (SNAP-tag, HaloTag7).
Chemically labeling the self-labeling tags with various fluorophores.
Spectrophotometrically analyzing the emission spectra of labeled luciferases.

Main Results:

Successfully engineered luciferases whose emission maxima shift to match the attached fluorophore.
Demonstrated tunable emission from blue to far-red wavelengths.
Validated the functionality of these tunable luciferases in reporter gene assays.

Conclusions:

Chemically labeled NanoLuc chimeras provide a flexible platform for tunable light emission.
These engineered luciferases offer significant advantages for multiplexed reporter gene studies, biosensor construction, and in vivo bioimaging.
The ability to tune emission color expands the toolkit for advanced biological research.