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Protein biosensors based on the principle of fluorescence resonance energy transfer for monitoring cellular dynamics

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Abstract

Genetically-coded, fluorescence resonance energy transfer (FRET) biosensors are widely used to study molecular events from single cells to whole organisms. They are unique among biosensors because of their spontaneous fluorescence and targeting specificity to both organelles and tissues. In this review, we discuss the theoretical basis of FRET with a focus on key parameters responsible for designing FRET biosensors that have the highest sensitivity. Next, we discuss recent applications that are grouped into four common biosensor design patterns—intermolecular FRET, intramolecular FRET, FRET from substrate cleavage and FRET using multiple colour fluorescent proteins. Lastly, we discuss recent progress in creating fluorescent proteins suitable for FRET purposes. Together these advances in the development of FRET biosensors are beginning to unravel the interconnected and intricate signalling processes as they are occurring in living cells and organisms.

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Acknowledgements

This work was supported by grants from the Canadian Foundation of Innovation (CFI) and the National Science and Engineering Research Council (NSERC).

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Authors and Affiliations

  1. Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ont., Canada, M5S 3G9

    Isaac T. Li, Elizabeth Pham & Kevin Truong

  2. Edward S. Rogers Sr. Department of Electrical and Computer Engineering, University of Toronto, 10 King’s College Circle, Toronto, Ont., Canada, M5S 3G4

    Isaac T. Li, Elizabeth Pham & Kevin Truong

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  1. Isaac T. Li
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  2. Elizabeth Pham
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Correspondence to Kevin Truong.

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Li, I.T., Pham, E. & Truong, K. Protein biosensors based on the principle of fluorescence resonance energy transfer for monitoring cellular dynamics. Biotechnol Lett 28, 1971–1982 (2006). https://doi.org/10.1007/s10529-006-9193-5

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  • DOI: https://doi.org/10.1007/s10529-006-9193-5

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