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Article Dans Une Revue Science Advances Année : 2020

Isothermal digital detection of microRNAs using background-free molecular circuit

Guillaume Gines
Kaori Nara
  • Fonction : Auteur
Valérie Taly
Yannick Rondelez

Résumé

MicroRNAs, a class of transcripts involved in the regulation of gene expression, are emerging as promising diseasespecific biomarkers accessible from tissues or bodily fluids. However, their accurate quantification from biological samples remains challenging. We report a sensitive and quantitative microRNA detection method using an isothermal amplification chemistry adapted to a droplet digital readout. Building on molecular programming concepts, we design a DNA circuit that converts, thresholds, amplifies, and reports the presence of a specific microRNA, down to the femtomolar concentration. Using a leak absorption mechanism, we were able to suppress nonspecific amplification, classically encountered in other exponential amplification reactions. As a result, we demonstrate that this isothermal amplification scheme is adapted to digital counting of microRNAs: By partitioning the reaction mixture into water-in-oil droplets, resulting in single microRNA encapsulation and amplification, the method provides absolute target quantification. The modularity of our approach enables to repurpose the assay for various microRNA sequences.
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Origine : Publication financée par une institution

Dates et versions

hal-03058004 , version 1 (04-01-2021)

Identifiants

Citer

Guillaume Gines, Roberta Menezes, Kaori Nara, Anne-Sophie Kirstetter, Valérie Taly, et al.. Isothermal digital detection of microRNAs using background-free molecular circuit. Science Advances , 2020, 6 (4), pp.eaay5952. ⟨10.1126/sciadv.aay5952⟩. ⟨hal-03058004⟩
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