Abstract
Mammalian cells ubiquitously release membrane-enclosed vesicles, known as extracellular vesicles. These particles carry a variety of molecules that reflect the status of their cells of origin, making them valuable sources for biomarker discovery. Furthermore, extracellular vesicles deliver their cargo locally and systemically to regulate biological processes, piquing interest in modulating extracellular vesicle biogenesis and developing extracellular vesicle-based therapies. Therefore, a thorough understanding of the extracellular vesicle life cycle, from biogenesis and trafficking to degradation, is essential for unlocking their full potential in biomarker identification and for the design of extracellular vesicle-based therapies. In this Review, we start by outlining the key steps in the extracellular vesicle life cycle and highlight remaining open questions. We then discuss the design and application of genetically encoded systems that can be applied to study extracellular vesicle biogenesis and fate. Finally, we highlight technical challenges that remain to be addressed in the engineering and application of genetically encoded systems to extracellular vesicle research.
Key points
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Extracellular vesicles are shed by cells and implicated in biological processes through diverse mechanisms.
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Specific and sensitive characterization of extracellular vesicles is fundamental for identifying extracellular vesicle-based biomarkers and developing therapies. However, their small size and close resemblance to other non-vesicular extracellular particles make their characterization challenging.
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Genetically encoded systems allow the high-specificity and high-sensitivity characterization of the extracellular vesicle life cycle, from biogenesis and trafficking to degradation.
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Genetic circuits can be incorporated for the long-term tracking of extracellular vesicle biogenesis and biodistribution in vivo.
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Caution must be applied when extrapolating knowledge of genetically engineered extracellular vesicles to their native counterparts, which requires complementary approaches.
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Acknowledgements
S.E.A. was funded by the European Research Council under the European Union’s Horizon 2020 research and innovation programme (EXPERT, grant agreement No. 825828), the European Research Council Consolidator Grant (DELIVER, grant agreement No. 101001374), the Swedish Foundation of Strategic Research (FormulaEx, grant agreement No. SM19-0007), the Swedish Cancer Society (project agreement No. 21-1762-Pj-01-H) and the Swedish Research Council (project agreement No. 4-258/2021). The authors acknowledge M. Mowoe for linguistic editing.
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W.Z. researched data for the draft and contributed to the writing and editing of this manuscript. S.R. and H.Z. contributed to the discussion of content, writing and editing of this manuscript. M.L.C. and G.v.N. contributed to reviewing and editing of the manuscript before submission. J.Z.N. made a substantial contribution to the discussion of content. S.E.A. contributed to the discussion and reviewed the manuscript before submission.
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Zheng, W., Roudi, S., Zhou, H. et al. Genetic tools for investigating the life cycle of extracellular vesicles. Nat Rev Bioeng 3, 505–520 (2025). https://doi.org/10.1038/s44222-025-00286-6
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DOI: https://doi.org/10.1038/s44222-025-00286-6
