Selective In Situ Analysis of Mature microRNAs in Extracellular Vesicles Using a DNA Cage‐Based Thermophoretic Assay

Author:

Zhao Shuai12,Zhang Shaohua3,Hu Huijun4,Cheng Yangchang12,Zou Kexuan5,Song Jie56,Deng Jinqi12,Li Lele12,Zhang Xiao‐Bing4,Ke Guoliang4,Sun Jiashu12ORCID

Affiliation:

1. Beijing Engineering Research Center for BioNanotechnology CAS Key Laboratory of Standardization and Measurement for Nanotechnology National Center for Nanoscience and Technology Beijing 100190 China

2. School of Future Technology University of Chinese Academy of Sciences Beijing 100049 China

3. Department of Oncology Senior Department of Oncology the Fifth Medical Center of PLA General Hospital Beijing 100071 China

4. Molecular Science and Biomedicine Laboratory State Key Laboratory of Chemo/Biosensing and Chemometrics College of Chemistry and Chemical Engineering Hunan University Changsha Hunan 410082 China

5. Institute of Nano Biomedicine and Engineering Department of Instrument Science and Engineering School of Electronic Information and Electrical Engineering Shanghai Jiao Tong University Shanghai 200240 China

6. Hangzhou Institute of Medicine Chinese Academy of Sciences Hangzhou Zhejiang 310022 China

Abstract

AbstractMature microRNAs (miRNAs) in extracellular vesicles (EVs) are involved in different stages of cancer progression, yet it remains challenging to precisely detect mature miRNAs in EVs due to the presence of interfering RNAs (such as longer precursor miRNAs, pre‐miRNAs) and the low abundance of tumor‐associated miRNAs. By leveraging the size‐selective ability of DNA cages and polyethylene glycol (PEG)‐enhanced thermophoretic accumulation of EVs, we devised a DNA cage‐based thermophoretic assay for highly sensitive, selective, and in situ detection of mature miRNAs in EVs with a low limit of detection (LoD) of 2.05 fM. Our assay can profile EV mature miRNAs directly in serum samples without the interference of pre‐miRNAs and the need for ultracentrifugation. A clinical study showed that EV miR‐21 or miR‐155 had an overall accuracy of 90 % for discrimination between breast cancer patients and healthy donors, which outperformed conventional molecular probes detecting both mature miRNAs and pre‐miRNAs. We envision that our assay can advance EV miRNA‐based diagnosis of cancer.

Funder

National Natural Science Foundation of China

National Key Research and Development Program of China

Chinese Academy of Sciences

Natural Science Foundation of Hunan Province

China Postdoctoral Science Foundation

Publisher

Wiley

Subject

General Medicine

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