Locked Nucleic Acid-Enhanced Entropy-Driven Amplifier Combined with Catalytic Hybridization Reaction-Based DNA Circuit for Dual Amplified Detection of Single Nucleotide Polymorphisms and Asymmetric Encryption of Gene Information

Abstract

Single-nucleotide polymorphisms (SNPs) play a pivotal role in investigations of disease-associated genes and in the genetic analysis of animal and plant varieties. Therefore, the detection of SNPs is essential for advancing biomedical diagnostics and therapeutics. Here, we report a locked nucleic acid (LNA)-enhanced dual signal amplification strategy for high-contrast detecting single-nucleotide polymorphisms (SNPs) in the KRAS_G12C gene. By integrating entropy-driven amplification with catalytic hybridization reaction, the proposed method achieves significant amplification of fluorescence and resonance Rayleigh scattering signals. The incorporation of LNA modification enhances the thermodynamic stability and reaction kinetics of the DNA computing circuit, resulting in superior sensitivity and specificity for SNPs detection. The method exhibits a low detection limit of 0.19 fM and a wide dynamic range from 1 fM to 0.1 nM for the KRAS_G12C gene. Compared to traditional DNA-based circuits, the LNA-modified system demonstrates enhanced discrimination of single-base mismatches and improved signal gain. Moreover, the proposed method was further demonstrated for its potential application in human serum samples. Impressively, this research not only presents a highly sensitive and selective platform for SNPs detection but also demonstrates its potential for molecular-level information encryption. The incorporation of LNA in dual signal amplification significantly elevates the intricacy and robustness of information encryption. Therefore, this study underscores the potential of DNA-based technologies to serve as a bridge between the era of biomedical research and the emerging Internet of things.