Target recognition initiated self-dissociation based DNA nanomachine for sensitive and accurate MicroRNA (miRNA) detection

Anal Biochem. 2023 Feb 1:662:115014. doi: 10.1016/j.ab.2022.115014. Epub 2022 Dec 6.

Abstract

As a valuable biomarker for various tumor, sensitive and reliable quantitative determination of microRNA (miRNA) is crucial for both disease diagnosis and cancer treatment. Herein, we depict a novel simple and sensitive miRNA detection approach by exploiting an elegantly designed target recognition initiated self-dissociation based DNA nanomachine. In this nanomachine, target recognition assists the formation of active DNAzyme secondary conformation, and the active DNAzyme generates a nicking site in H probe, initiating the self-assembly of H probe. With the reflexed sequences as primer, dual signal recycles are formed under the cooperation of DNA polymerase and Nb.BbvCI. Eventually, the method exhibits a high sensitivity with the limit of detection as low as 12 fM. In addition, the method is also demonstrated with a high selectivity that can distinguish one mismatched base pair. We believe the established approach can be a robust tool for the early-diagnosis of a variety of cancers and also in anticancer drug development.

Keywords: EXPAR; Nb.BbvCI; Switchable nanomachine; miRNA.

MeSH terms

  • Biosensing Techniques* / methods
  • DNA
  • DNA, Catalytic*
  • Limit of Detection
  • MicroRNAs* / genetics
  • Nucleic Acid Amplification Techniques / methods

Substances

  • MicroRNAs
  • DNA, Catalytic
  • DNA