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Related Experiment Video

Updated: Jan 16, 2026

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Programmable DNAzyme switch integrated with low-background cross-reaction for sensitive and selective SNP

Jing Ye1, Chunyan Liu2, Ximiao Yang1

  • 1Key Laboratory of Soybean Molecular Design Breeding, National Key Laboratory of Black Soils Conservation and Utilization, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Changchun, 130102, China.

Biosensors & Bioelectronics
|October 3, 2025
PubMed
Summary

This study introduces a novel DNAzyme strategy for highly selective single-nucleotide polymorphism (SNP) detection. The innovative approach enhances precision genotyping and molecular diagnostics, overcoming limitations of current methods.

Keywords:
BiosensorDNAzymeElectrochemicalEnzyme-freeSingle nucleotide polymorphismsSolid–liquid phase amplification

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Area of Science:

  • Biotechnology and Molecular Diagnostics
  • Genetics and Genomics
  • Biosensing Technologies

Background:

  • Single-nucleotide polymorphism (SNP) detection is crucial for disease screening, personalized medicine, and crop improvement.
  • DNAzymes offer sequence programmability and catalytic activity but suffer from limited selectivity and high nonspecific reactivity for precise SNP discrimination.
  • Existing enzyme-free methods often face stoichiometric limitations, impacting target selectivity.

Purpose of the Study:

  • To overcome the limitations of current DNAzyme-based SNP detection systems, particularly concerning sequence selectivity and nonspecific reactivity.
  • To develop a highly sensitive and selective enzyme-free platform for SNP recognition and genotyping.
  • To enhance analytical sensitivity and address the trade-off between selectivity and sensitivity in biosensing.

Main Methods:

  • Redesigned the DNAzyme catalytic core to create a competitive molecular switch with an 'activation-silencing' mechanism for single-base specificity.
  • Implemented a protein-enzyme-free strategy for SNP recognition, breaking conventional stoichiometric paradigms.
  • Introduced a solid-liquid phase cross-reaction mechanism and a cascade system with electrochemical biosensing to boost analytical sensitivity.

Main Results:

  • Achieved highly selective SNP recognition with enhanced target selectivity compared to conventional methods.
  • Demonstrated a significantly improved detection limit of 11.3 aM for SNPs, surpassing conventional sensors (370 aM).
  • Showcased high consistency in genotyping soybean variants, validating the platform's practical applicability.

Conclusions:

  • The developed strategy provides an innovative, enzyme-free, and scalable platform for SNP sensing and signal regulation.
  • This work offers new concepts for precision genotyping and molecular diagnostics by improving single-base recognition and signal transduction.
  • The approach effectively addresses the bottleneck of single-base specificity and enhances detection sensitivity, paving the way for advanced diagnostic tools.