High-Fidelity Determination and Tracing of Small Extracellular Vesicle Cargoes
High-Fidelity Determination and Tracing of Small Extracellular Vesicle Cargoes
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小细胞外囊泡货物的高保真测定和示踪
DOI：
10.1002/smll.202002800
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发表时间：
2020-09-02
期刊：
影响因子：
13.3
通讯作者：
Luo, Yang
中科院分区：
文献类型：
--
作者：
Chen, Xiaohui;Jia, Mei;Luo, Yang
直接追踪小细胞外囊泡（sEV）货物对于阐明涉及细胞间通讯的机制具有前所未有的重要性。然而，由于难以将分子探针运输到完整的sev中，因此尚未实现对sev分子载体的高保真测定。本文描述了一种荧光细胞内引导的发夹-四面体（fLIGHT）纳米探针，用于直接原位可视化sEV microrna。结合DNA折纸和单核苷酸识别的非破坏性sEV穿透以及使用发夹探针的无水洗荧光读取的优势，该方法可以实现sEV的microRNA的高保真荧光可视化，而无需提取或泄漏RNA，展示了sEV货物现场追踪的潜力。这一策略为建立通用的分子检测和标记平台开辟了一条道路，可以促进sev衍生的基础生物学研究和分子诊断。
Direct tracing of small extracellular vesicle (sEV) cargoes holds unprecedented importance for elucidating the mechanisms involved in intercellular communication. However, high-fidelity determination of sEVs' molecular cargoes in situ has yet to be achieved due to the difficulty in transporting molecular probes into intact sEVs. Herein, a fLuorescent Intracellular-Guided Hairpin-Tetrahedron (fLIGHT) nanoprobe is described for direct visualization of sEV microRNAs in situ. Integrating the advantages of nondestructive sEV penetration via DNA origami and single-nucleotide discrimination as well as wash-free fluorescence readout using a hairpin probe, the proposed approach enables high-fidelity fluorescence visualization of sEVs' microRNA without RNA extraction or leakage, demonstrating the potential of on-site tracing of sEV cargoes. This strategy opens an avenue to establishing universal molecular detection and labeling platforms that can facilitate both sEV-derived fundamental biological studies and molecular diagnostics.