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DNA Zipper Mediated Membrane Fusion for Rapid Exosomal MiRNA Detection

Authors :
Miao Xie
Fujun Wang
Jiapei Yang
Yuanyuan Guo
Fei Ding
Xinmiao Lu
Yangyang Huang
Yimeng Li
Xinyuan Zhu
Chuan Zhang
Source :
Analytical Chemistry. 94:13043-13051
Publication Year :
2022
Publisher :
American Chemical Society (ACS), 2022.

Abstract

Accurate and reliable detection of exosomal miRNA can serve as a promising method for early diagnosis of disease and evaluation of therapeutic effects. However, current exosomal miRNA detection methods commonly involve exosome enrichment, containing RNA extraction, and qRT-PCR based quantification, which are expensive and time-consuming. Herein, we develop a DNA zipper-mediated membrane fusion approach for rapid exosomal miRNA detection and cancer diagnosis. First, a lipid vesicle probe containing miR21-targeting molecular beacons (MBs) is constructed and further loaded with zipper DNA constructs (ZDCs) on its surface. Meanwhile, complementary zipper DNA constructs (cZDCs) are introduced on the exosome of interest. Upon mixing them together, zipping between ZDC and cZDC induces the membrane fusion of exosomes and vesicle probes, triggering the recognition of exosomal miR21 by contained MBs and fluorescence emission that can be conveniently detected within 30 min. Importantly, with the assistance of flow cytometry, miR21-overexpressed tumor exosomes derived from either cell culture medium or clinical patient serums can be distinguished from exosomes secreted from normal cells. This approach provides a convenient way to accurately detect the exosomal miRNA, which may hold great potential in liquid biopsy for early cancer diagnosis and monitoring the therapeutic effects during the treatments.

Details

ISSN :
15206882 and 00032700
Volume :
94
Database :
OpenAIRE
Journal :
Analytical Chemistry
Accession number :
edsair.doi.dedup.....5dde0b6d39cdfed5d1209bc15befa98c
Full Text :
https://doi.org/10.1021/acs.analchem.2c01980