Li Po, Lei Xueying, Niu Xiaoying, Tian Wen, Li Zhehuang, Yu Songcheng, Zhang Peng
Department of Orthopedic and Soft Tissue, The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, Zhengzhou, 450008, China.
College of Public Health, Zhengzhou University, Zhengzhou, 450001, China.
Synth Syst Biotechnol. 2025 Jul 5;10(4):1208-1214. doi: 10.1016/j.synbio.2025.07.002. eCollection 2025 Dec.
MicroRNA (miRNA) is promising candidate for non-invasive diagnostic biomarker. Conventional CRISPR/Cas12a-based miRNA detection systems are constrained by reliance on reverse transcription, nucleic acid pre-amplification and costly fluorescently labeled reporters which introduce chemical modification complexity and background noise. To address these limitations, we herein developed a multifunctional switch that integrated target recognition, CRISPR/Cas12a system activation, intrinsic fluorescence signaling, and autonomous signal amplification within a single molecular architecture. As a proof of concept, this switch enabled a label-free CRISPR/Cas12a biosensing for miR-21 detection with a detection limit of 4.8 nM and robust performance in accuracy, precision, and selectivity. This proposed label-free CRISPR/Cas12a platform could be applied for real sample and is a promising candidate for point-of-care miRNA detection.
微小RNA（miRNA）是无创诊断生物标志物的理想候选者。传统的基于CRISPR/Cas12a的miRNA检测系统受到依赖逆转录、核酸预扩增以及昂贵的荧光标记报告分子的限制，这些因素会带来化学修饰复杂性和背景噪声。为了解决这些限制，我们在此开发了一种多功能开关，该开关在单个分子结构中集成了靶标识别、CRISPR/Cas12a系统激活、固有荧光信号传导和自主信号放大功能。作为概念验证，这种开关实现了用于miR-21检测的无标记CRISPR/Cas12a生物传感，检测限为4.8 nM，在准确性、精密度和选择性方面具有强大性能。所提出的无标记CRISPR/Cas12a平台可应用于实际样品，是即时检测miRNA的有前景的候选者。
