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Nucleic acid detection using BRET-beacons based on bioluminescent protein-DNA hybrids
Wouter Engelen1, Kayleigh M van de Wiel1, Lenny H H Meijer1
1Laboratory of Chemical Biology and Institute for Complex Molecular Systems Eindhoven, University of Technology, Den Dolech 2, 5600 MB, Eindhoven, The Netherlands. m.merkx@tue.nl.
Chemical Communications (Cambridge, England)
|February 21, 2017
Summary
New bioluminescent molecular beacons use NanoLuc and Cy3 for sensitive nucleic acid detection. These BRET-beacons overcome limitations of traditional methods, enabling detection in complex samples.
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Classical molecular beacons face challenges with background fluorescence and light scattering.
- These limitations hinder sensitive detection of nucleic acids in complex biological or environmental samples.
Purpose of the Study:
- To develop novel bioluminescent molecular beacons with improved sensitivity and reduced background noise.
- To enable direct detection of nucleic acids in complex media using a modular design.
Main Methods:
- Utilized a modular design approach for bioluminescent molecular beacons.
- Employed bioluminescence resonance energy transfer (BRET) between a bright luciferase (NanoLuc) and a Cy3 acceptor fluorophore.
- Developed BRET-beacons for nucleic acid detection.
Main Results:
- Successfully developed BRET-beacons leveraging NanoLuc and Cy3.
- Demonstrated the ability to detect low picomolar (pM) concentrations of nucleic acids.
- Showcased direct detection of nucleic acids within complex media, overcoming background interference.
Conclusions:
- Bioluminescent BRET-beacons offer a sensitive and robust alternative to traditional molecular beacons.
- The modular design and BRET principle allow for reliable nucleic acid quantification in challenging sample types.
- This technology holds promise for various applications in molecular diagnostics and research.
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