Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8946491 | Biomaterials | 2018 | 31 Pages |
Abstract
Detection of specific extracellular RNAs has been developed for non-invasive cancer diagnosis. However, accurate and efficient identification of RNAs with single-point mutation in cancer cells-derived extracellular vesicles (EVs) is challenging. Herein, we present a unique overhang molecular beacon with internal dye (Ohi-MB) with a stable hairpin structure, fast hybridization kinetics and single mismatch specificity. Ohi-MBs are encapsulated in cationic lipoplex nanoparticles (CLNs) that are tethered on a gold coated glass slide as a chip, which can capture circulating EVs and detect encapsulated target RNAs in-situ in a single step. The capability of detection of single-point mutation by CLN-Ohi-MB is demonstrated in artificial EVs and cancer cells. This CLN-Ohi-MB biochip could quantify single-point mutations in KRAS mRNA (G12C, G12D, G12V) in pancreatic cancer cell-derived EVs and single-point mutations in EGFR mRNA (L858R and T790M) in lung cancer cell-derived EVs with high specificity, not achievable by conventional molecular probes. We show that CLN-Ohi-MB biochip could selectively and sensitively identify single-point mutations in KRAS mRNA in human serum EVs, distinguishing pancreatic cancer patients with different mutations.
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Physical Sciences and Engineering
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Authors
Jiaming Hu, Kwang Joo Kwak, Junfeng Shi, Bohao Yu, Yan Sheng, Ly James Lee,