| Autor: |
Zhang S; Department of Hepatobiliary, Taizhou Central Hospital, Taizhou University, Taizhou, Zhejiang 318000, China.; School of Pharmaceutical and Chemical Engineering, Taizhou University, Jiaojiang, Taizhou 318000, Zhejiang, China., Shao H; Zhijiang College of Zhejiang University of Technology, Shaoxing, Zhejiang 312000, China., Shi W; School of Pharmaceutical and Chemical Engineering, Taizhou University, Jiaojiang, Taizhou 318000, Zhejiang, China., Li KB; School of Pharmaceutical and Chemical Engineering, Taizhou University, Jiaojiang, Taizhou 318000, Zhejiang, China., You N; College of Petrochemical Engineering, Liaoning Petrochemical University, Fushun 113001, China., Han DM; Department of Hepatobiliary, Taizhou Central Hospital, Taizhou University, Taizhou, Zhejiang 318000, China.; School of Pharmaceutical and Chemical Engineering, Taizhou University, Jiaojiang, Taizhou 318000, Zhejiang, China., Mo J; Department of Hepatobiliary, Taizhou Central Hospital, Taizhou University, Taizhou, Zhejiang 318000, China. |
| Abstrakt: |
The detection of DNA methyltransferase (MTase) was crucial for understanding gene expression regulation, cancer mechanisms, and various biological processes, contributing significantly to disease diagnosis and drug development. Herein, a nanopore sensor based on cascaded signal amplification of DNA walker and autocatalytic hybridization reaction (AHR) was developed for the ultrasensitive determination of various MTases. In the presence of Dam MTase, the hairpin structure H D underwent methylation and cleavage by DpnI endonuclease, forming T-DNA fragments. These T-DNA fragments were used to activate the DNA walker, which moved across the surface of magnetic beads step by step, generating a large quantity of initiator I by cleaving the substrate. The initiator I subsequently activated the AHR. The AHR included a hybridization chain reaction (HCR) amplifier and a catalytic hairpin assembly (CHA) convertor. The HCR amplifier generated multiple novel CHA triggers, which activated the CHA convertor. This, in turn, stimulated the HCR amplifier, creating an AHR circuit that resulted in the formation of numerous DNA nanowires. These DNA nanowires were adsorbed onto the G4-PAMAM-modified nanopore surface under the influence of an electric field, thereby altering the surface charge of the nanopore and changing the ionic rectification curve. The detection limit of the Dam MTase nanopore sensor reached 0.0002 U/mL. By modification of the recognition sites of the probes, this nanopore system could also be used for the detection of M.SssI MTase. Moreover, a four-input parallel concatenated logic circuit (AND//INHIBIT-OR) had been constructed and applied for the multivariate detection of Dam MTase and M.SssI MTase, presenting a novel conceptual model for advancing the construction of nanopore logic gate systems and their applications in biosensing. |
