3D matrixed DNA self-nanocatalyzer as electrochemical sensitizers for ultrasensitive investigation of DNA 5-methylcytosine

Abstract

DNA methylation plays an important role in a variety of human diseases. Thus, accurately analyze 5-methylcytosine in different DNA segments is of great significance. Herein, we proposed a novel 3D matrixed DNA self-nanocatalyzer via gold nanoparticles (AuNPs) supporting DNA self-hybridization with hemin as biomimetic enzyme and methylene blue (MB) as electrochemical mediator, which was employed as an efficient electrochemical sensitizer for the ultrasensitive bioassay of DNA 5-methylcytosine. Meanwhile, the AuNPs, graphitic carbon nitride (g-C3N4) and reduced graphene oxide (rGO) was prepared as AuNPs/g-C3N4@rGO nanocomposites to coat on the electrode surface to immobilize the capture hairpin DNA (CH). In the presence of target DNA with 5-methylcytosine, the target DNA could hybridize with CH via the hyperstable triple-helix formation. Based on the specific biorecognition between biotin and streptavidin and immune recognition between anti-5-methylcytosine antibodies and 5-methylcytosine sites on the target DNA, the 3D matrixed DNA self-nanocatalyzer could be captured onto the electrode surface to generate an amplified electrochemical signal related to the concentration of 5-methylcytosine. Under the optimal conditions, the proposed strategy performed a linear range from 10−17 M to 10−8 M with a detection limit of 8.6 aM. Remarkably, this strategy could be expanded easily to various biomarkers, including protein, DNA, phosphorylation and glycosylation, providing a promising strategy for clinical diagnosis and mechanism investigation of various diseases.