<p>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.</p>