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A general method for the development of multicolor biosensors with large dynamic ranges
Lars Hellweg1, Anna Edenhofer1, Lucas Barck1
1Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany.
Nature Chemical Biology
|June 8, 2023
Summary
Researchers developed new fluorescent biosensors using Förster resonance energy transfer (FRET) pairs. These biosensors offer significantly improved dynamic ranges for studying cellular processes like calcium, ATP, and NAD+ levels.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Fluorescent biosensors are crucial for studying cell physiology with high spatiotemporal resolution.
- Existing biosensors often have limited dynamic ranges, hindering detailed cellular analysis.
Purpose of the Study:
- To introduce a novel family of Förster resonance energy transfer (FRET) pairs for enhanced biosensor development.
- To create highly sensitive and tunable biosensors for key cellular analytes.
Main Methods:
- Designed FRET pairs based on reversible interactions between fluorescent proteins and HaloTag.
- Developed biosensors for calcium, adenosine triphosphate (ATP), and nicotinamide adenine dinucleotide (NAD+).
- Modified biosensors for readout via fluorescence intensity, lifetime, or bioluminescence.
Main Results:
- Achieved near-quantitative FRET efficiencies with the designed FRET pairs.
- Developed biosensors for calcium, ATP, and NAD+ with unprecedented dynamic ranges.
- Demonstrated tunable colors for simultaneous monitoring of NAD+ in different cellular compartments.
Conclusions:
- Introduced a new concept for developing highly sensitive and tunable biosensors.
- The designed FRET pairs significantly advance the capabilities of fluorescent biosensors.
- Enabled precise monitoring of cellular analytes under various conditions, including genotoxic stress.
