Entropy-driven self-assembled enzyme-DNA nanomatrix cascade DNAzyme-CRISPR/cas system for multiplexed enhancement of self-powered sensing platform for protein detection

• Published in: International journal of biological macromolecules 2024-12, Vol.282 (Pt 6), p.137536, Article 137536
• Main Authors: Ma, Yunzhi, Ya, Yu, Wu, Ye-Yu, Yan, Jun, Huang, Ke-Jing, Tan, Xue-Cai, Duan, Yun
• Format: Article
• Language: English
• Subjects: Aptamers, Nucleotide - chemistry; Biosensing Techniques - methods; C-MoS2 hollow nanorods; CRISPR-Cas Systems; CRISPR/Cas system; DNA - chemistry; DNA - genetics; DNA - metabolism; DNA, Catalytic - chemistry; DNA, Catalytic - metabolism; Entropy; Entropy-driven self-assembly enzyme; Glucose Oxidase - chemistry; Glucose Oxidase - metabolism; Nanotubes - chemistry; Thrombin - analysis; Thrombin - metabolism
• ISSN: 0141-8130; 1879-0003; 1879-0003
• DOI: 10.1016/j.ijbiomac.2024.137536

Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated Proteins (CRISPR/Cas) system can accurately identify and cleave target DNA sequences, while the effective combination of DNA nanomatrix and entropy-driven self-assembled enzymes can significantly enhance the sensitivity, stability, and diversified functionality of sensors through highly ordered molecular arrangement and spontaneous efficient assembly processes. Herein, a carbon-encapsulated MoS2 hollow nanorod (C-MoS2) with excellent conductivity and multiple active sites is used to construct bioanode of biofuel cell by integrating it with an entropy-driven self-assembled enzyme-DNA nanomatrix cascade DNAzyme-CRISPR/Cas system. When thrombin binds aptamer, it exposes the trigger strand on the anode, initiating chain displacement. This activates DNAzyme, triggering a cascade reaction that cleaves and releases the probe strand. The probe then binds CrRNA, forming a multimeric complex with Cas protein.When non-specific cleavage of the single strand occurs, it leads to the release of glucose oxidase (GOD) from the DNA matrix and causes a significant decrease in the value of the system's open-circuit voltage (EOCV). The EOCV values show a good negative correlation with the concentration of thrombin (TB) in the range of 0.00001–100 nM, achieving a limit of detection of 3.55 fM (S/N = 3). •Entropy-driven self-assembly of enzyme-DNA nanomatrices enables fine-tuning of the - structures of DNA nanostructures.•DNAzyme-CRISPR/Cas system, when combined with C-MoS2, can accurately recognize targets and amplify the output signal.•The integration of capacitors and smartphones in a detection platform enables sensitive and rapid detection of the target.