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Enzyme-free amplified detection of cellular microRNA by light-harvesting fluorescent nanoparticle probes.
- Source :
-
Biosensors & Bioelectronics . May2021, Vol. 179, pN.PAG-N.PAG. 1p. - Publication Year :
- 2021
-
Abstract
- Detection of cellular microRNA biomarkers is an emerging powerful tool in cancer diagnostics. Currently, it requires multistep tedious protocols based on molecular amplification of the RNA target, e.g. RT-qPCR. Here, we developed a one-step enzyme-free method for microRNA detection in cellular extracts based on light-harvesting nanoparticle (nanoantenna) biosensors. They amplify the fluorescence signal by effective Förster resonance energy transfer (FRET) from ultrabright dye-loaded polymeric nanoparticle to a single acceptor and thus convert recognition of one microRNA copy (through nucleic acid strand displacement) into a response of >400 dyes. The developed nanoprobes of 17–19 nm diameter for four microRNAs (miR-21, let-7f, miR-222 and miR-30a) exhibit outstanding brightness (up to 3.8 × 107 M−1cm−1) and ratiometric sequence-specific response to microRNA with the limit of detection (LOD) down to 1.3 pM (21 amol), equivalent to 24 RT-qPCR cycles. They enable quantitative detection of the four microRNAs in RNA extracts from five cancerous cell lines (human breast cancer - T47D and MCF7, head and neck cancer - CAL33 and glioblastoma - LNZ308 and U373) and two non-cancerous ones (Hek293 and MCF10A), in agreement with RT-qPCR. The results confirmed that let-7f and especially miR-21 are systematically overexpressed in all studied cancerous cell lines. These nanoparticle biosensors are compatible with low-cost portable fluorometers and small detection volumes (11 amol LOD), opening a route to rapid point-of-care cancer diagnostics. • Fluorescent nanoprobes for microRNA detection in cell extracts are developed. • They amplify signal up to 130-fold by light-harvesting from ~400 FRET donor dyes. • They enable ratiometric analysis of microRNA with limit of detection down to 1.3 pM. • Biosensors for four microRNA are validated in 5 cancer and 2 healthy cell lines. • They are compatible with low-cost portable fluorometers and small detection volumes. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09565663
- Volume :
- 179
- Database :
- Academic Search Index
- Journal :
- Biosensors & Bioelectronics
- Publication Type :
- Academic Journal
- Accession number :
- 149087877
- Full Text :
- https://doi.org/10.1016/j.bios.2021.113084