Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Department of Pharmaceutical Sciences, University at Buffalo, USA.
J Control Release. 2024 Oct;374:349-368. doi: 10.1016/j.jconrel.2024.07.043. Epub 2024 Aug 22.
Extracellular vesicles (EVs), or exosomes, play important roles in physiological and pathological cellular communication and have gained substantial traction as biological drug carriers. EVs contain both short and long non-coding RNAs that regulate gene expression and epigenetic processes. To fully capitalize on the potential of EVs as drug carriers, it is important to study and understand the intricacies of EV function and EV RNA-based communication. Here we developed a genetically encodable RNA-based biomaterial, termed EXO-Probe, for tracking EV RNAs. The EXO-Probe comprises an EV-loading RNA sequence (EXO-Code), fused to a fluorogenic RNA Mango aptamer for RNA imaging. This fusion construct allowed the visualization and tracking of EV RNA and colocalization with markers of multivesicular bodies; imaging RNA within EVs, and non-destructive quantification of EVs. Overall, the new RNA-based biomaterial provides a useful and versatile means to interrogate the role of EVs in cellular communication via RNA trafficking to EVs and to study cellular sorting decisions. The system will also help lay the foundation to further improve the therapeutic efficacy of EVs as drug carriers.
细胞外囊泡 (EVs)，或外泌体，在生理和病理细胞通讯中发挥着重要作用，并作为生物药物载体获得了广泛关注。EVs 中含有短链和长链非编码 RNA，它们调节基因表达和表观遗传过程。为了充分利用 EVs 作为药物载体的潜力，研究和了解 EV 功能和基于 EV RNA 的通讯的复杂性非常重要。在这里，我们开发了一种基于遗传编码的 RNA 生物材料，称为 EXO-Probe，用于跟踪 EV RNA。EXO-Probe 由 EV 加载 RNA 序列 (EXO-Code) 与荧光 RNA Mango 适体融合而成，用于 RNA 成像。这种融合结构允许可视化和跟踪 EV RNA，并与多泡体标志物共定位；在 EV 内成像 RNA，并对 EV 进行非破坏性定量。总的来说，这种新的基于 RNA 的生物材料为通过 RNA 向 EV 的运输来研究 EV 在细胞通讯中的作用以及研究细胞分选决策提供了一种有用且多功能的方法。该系统还将有助于为进一步提高 EV 作为药物载体的治疗效果奠定基础。
