Monitoring VEGF mRNA and imaging in living cells in vitro using rGO-based dual fluorescent signal amplification platform

Abstract

VEGF mRNA, as an important biomarker for disease diagnosis and therapeutics, has received extensive attention. However, how to monitor its mRNA levels rapidly and sensitively remains a challenge. Herein, a strategy was designed for facile and efficient detection of VEGF mRNA and imaging in living cells using a collaborative system of a fluorophore-labeled single-stranded probe (P), reduced graphene oxide (rGO) and double-specific nuclease (DSN). The combination of strong fluorophore-quenching ability of rGO with DSN assisted signal amplification contributes to the superior sensitivity of the assay for VEGF mRNA, which was reflected by the lower limit of mRNA detection of 100 fM obtained using dual signal amplification manner. Furthermore, the developed sensor was directly used for intracellular mRNA imaging in vitro without the assistance of transfection reagent. In summary, the simple, ultra-sensitive and cost-effective mRNA assay system, which provided a general analysis strategy for other mRNAs assay by replacing the sequence of the probe, is hopeful for applying on the clinical diagnosis and therapy.