An Ultrasensitive and Efficient microRNA Nanosensor Empowered by the CRISPR/Cas Confined in a Nanopore

作者：Bing Wang, Yi‐Tong Xu, Tianyang Zhang, Haiyan Wang, Xian Zhang, Zeng‐Qiang Wu, Weiwei Zhao, Hong‐Yuan Chen, Jing‐Juan Xu · 发表于：Nano Letters · 年份：2023 · DOI：10.1021/acs.nanolett.3c03723 · 被引用次数：42 · 研究领域：Advanced biosensing and bioanalysis techniques、CRISPR and Genetic Engineering、Nanopore and Nanochannel Transport Studies

This work presents a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas-nanopipette nano-electrochemistry (Cas = CRISPR-associated proteins) capable of ultrasensitive microRNA detection. Nanoconfinement of the CRISPR/Cas13a within a nanopipette leads to a high catalytic efficacy of ca. 169 times higher than that in bulk electrolyte, contributing to the amplified electrochemical responses. CRISPR/Cas13a-enabled detection of representative microRNA-25 achieves a low limit of detection down to 10 aM. Practical application of this method is further demonstrated for single-cell and real human serum detection. Its general applicability is validated by addressing microRNA-141 and the SARS-CoV-2 RNA gene fragment. This work introduces a new CRISPR/Cas-empowered nanotechnology for ultrasensitive nano-electrochemistry and bioanalysis.