Abstract
Current methods for characterizing the biodistribution of extracellular vesicles (EVs) are not sensitive enough to track EVs in vivo, despite significant advances over the past decade. Commonly used lipophilic fluorescent dyes are convenient, but lack specificity and yield inaccurate spatiotemporal images in the long-term tracking of EVs. In contrast, protein-based fluorescent or bioluminescent EV reporters have more accurately revealed their distribution in cells and mouse models. Here, we describe a red-shifted bioluminescence resonance energy transfer (BRET) EV reporter, PalmReNL, to analyze the trafficking of small EVs (<200 nm; sEVs) and medium/large EVs (>200 nm; m/lEVs) in mice. Its advantages are that (i) background signals in bioluminescence imaging (BLI) are negligible and (ii) the photons PalmReNL emits have spectral wavelengths longer than 600 nm and can more efficiently penetrate tissues than reporters emitting shorter wavelength light.
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Acknowledgments
We thank the Institute for Quantitative Health Science and Engineering (IQ) for providing facilities and resources for executing this work. This work was supported by start-up funds from MSU to MK.
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Perez, G.I., Bachmann, M.H., Kanada, M. (2023). In Vivo Analysis of Heterogeneous Extracellular Vesicles Using a Red-Shifted Bioluminescence Resonance Energy Transfer Reporter Protein. In: Vainio, S. (eds) Cell-Secreted Vesicles. Methods in Molecular Biology, vol 2668. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3203-1_3
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DOI: https://doi.org/10.1007/978-1-0716-3203-1_3
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