Co-freezing localized CRISPR-Cas12a system enables rapid and sensitive nucleic acid analysis

作者：Lifeng Zhang, Shihua Luo, Wenbin Li, Wanting Su, Siting Chen, Chunchen Liu, Weilun Pan, Bo Situ, Lei Zheng, Ling Li, Xiaohui Yan, Zhang Ye · 发表于：Journal of Nanobiotechnology · 年份：2024 · DOI：10.1186/s12951-024-02831-8 · 被引用次数：16 · 研究领域：CRISPR and Genetic Engineering、Advanced biosensing and bioanalysis techniques、Biosensors and Analytical Detection

Rapid and sensitive nucleic acid detection is vital in disease diagnosis and therapeutic assessment. Herein, we propose a co-freezing localized CRISPR-Cas12a (CL-Cas12a) strategy for sensitive nucleic acid detection. The CL-Cas12a was obtained through a 15-minute co-freezing process, allowing the Cas12a/crRNA complex and hairpin reporter confined on the AuNPs surface with high load efficiency, for rapid sensing of nucleic acid with superior performance to other localized Cas12a strategies. This CL-Cas12a based platform could quantitatively detect targets down to 98 aM in 30 min with excellent specificity. Furthermore, the CL-Cas12a successful applied to detect human papillomavirus infection and human lung cancer-associated single-nucleotide mutations. We also achieved powerful signal amplification for imaging Survivin mRNA in living cells. These findings highlight the potential of CL-Cas12a as an effective tool for nucleic acid diagnostics and disease monitoring.