Institute of Molecular Biotechnology, Department of Biotechnology, BOKU University, Vienna, Austria.
Ludwig Boltzmann Institute for Traumatology, The Research Center in Cooperation with AUVA, Vienna, Austria.
J Extracell Vesicles. 2024 Oct;13(10):e12523. doi: 10.1002/jev2.12523.
Extracellular vesicles (EVs) are lipid nanoparticles and play an important role in cell-cell communications, making them potential therapeutic agents and allowing to engineer for targeted drug delivery. The expanding applications of EVs in next generation medicine is still limited by existing tools for scaling standardized EV production, single EV tracing and analytics, and thus provide only a snapshot of tissue-specific EV cargo information. Here, we present the Snorkel-tag, for which we have genetically fused the EV surface marker protein CD81, to a series of tags with an additional transmembrane domain to be displayed on the EV surface, resembling a snorkel. This system enables the affinity purification of EVs from complex matrices in a non-destructive form while maintaining EV characteristics in terms of surface protein profiles, associated miRNA patterns and uptake into a model cell line. Therefore, we consider the Snorkel-tag to be a widely applicable tool in EV research, allowing for efficient preparation of EV standards and reference materials, or dissecting EVs with different surface markers when fusing to other tetraspanins in vitro or in vivo.
细胞外囊泡（EVs）是脂质纳米颗粒，在细胞间通讯中发挥重要作用，使它们成为有潜力的治疗药物，并可用于靶向药物递送的工程设计。EV 在下一代医学中的应用不断扩大，但受到现有标准化 EV 生产、单个 EV 追踪和分析工具的限制，因此只能提供组织特异性 EV 货物信息的快照。在这里，我们提出了 Snorkel-tag，我们将 EV 表面标记蛋白 CD81 与一系列带有额外跨膜结构域的标签融合，使其在 EV 表面上呈现出类似于通气管的结构。该系统可在非破坏性的方式下从复杂基质中亲和纯化 EV，同时保持 EV 特征，包括表面蛋白谱、相关 miRNA 模式和被模型细胞系摄取。因此，我们认为 Snorkel-tag 是 EV 研究中一种广泛适用的工具，可用于高效制备 EV 标准品和参考物质，或在体外或体内融合到其他四跨膜蛋白时对具有不同表面标记的 EV 进行剖析。
