Enzyme-free amplified detection of cellular microRNA by light-harvesting fluorescent nanoparticle probes.
Fiche publication
Date publication
février 2021
Journal
Biosensors & bioelectronics
Auteurs
Membres identifiés du Cancéropôle Est :
Dr MARTIN Sophie, Dr KLYMCHENKO Andrey
Tous les auteurs :
Egloff S, Melnychuk N, Reisch A, Martin S, Klymchenko AS
Lien Pubmed
Résumé
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 × 10 Mcm) 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.
Mots clés
Cancer biomarkers, Cellular extracts, Light-harvesting nanoantenna, MicroRNA detection, Nanoparticle biosensors, Point-of-care diagnostics
Référence
Biosens Bioelectron. 2021 Feb 10;179:113084