CRISPR-Responsive RCA-Based DNA Hydrogel Biosensing Platform with Customizable Signal Output for Rapid and Sensitive Nucleic Acid Detection

• Published in: Analytical chemistry (Washington) Vol. 96; no. 40; pp. 15998 - 16006
• Main Authors: Zhang, Yan, Wang, Weiwei, Zhou, Xinxi, Lin, Haonan, Zhu, Xiaotong, Lou, Yongliang, Zheng, Laibao
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 08.10.2024
• Subjects: Adaptability; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Biosensing Techniques - methods; Biosensors; Colorimetry; CRISPR; CRISPR-Associated Proteins; CRISPR-Cas Systems - genetics; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA - genetics; Drug resistance; Electrochemical Techniques; Electrochemistry; Endodeoxyribonucleases; Gold - chemistry; Humans; Hydrogels; Hydrogels - chemistry; Limit of Detection; MecA protein; Metal Nanoparticles - chemistry; Methicillin; Methicillin-Resistant Staphylococcus aureus - genetics; Methicillin-Resistant Staphylococcus aureus - isolation & purification; Methylene blue; Nanoparticles; Nucleic Acid Amplification Techniques; Nucleic acids; Penicillin-Binding Proteins; Target detection
• ISSN: 0003-2700; 1520-6882; 1520-6882
• DOI: 10.1021/acs.analchem.4c03450

Current nucleic acid-responsive DNA hydrogels face significant challenges, such as the requirement for high target concentrations, frequent redesigns, and increased costs, which limit their practical applications in biosensing. To address these issues, we developed a novel biosensing platform integrating a CRISPR/Cas12a system into an RCA-based DNA hydrogel. The hydrogel used in the platform could preencapsulate diverse signal molecules comprising GelRed, methylene blue, and gold nanoparticles, which were released upon Cas12a-mediated cleavage. This design enabled customizable signal output, including fluorescence, electrochemistry, and colorimetry, thereby ensuring the platform’s adaptability to various detection scenarios. Our platform was highly specific for methicillin-resistant Staphylococcus aureus, with a mecA gene detection limit of 10 copies/μL, and provided fast and accurate results within 2 h for clinical samples. Hence, based on these advantages, the proposed biosensing platform exhibits promising application prospects in the field of nucleic acid detection.