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

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An Ultrasensitive and Efficient microRNA Nanosensor Empowered by the CRISPR/Cas Confined in a Nanopore
Nano Letters ( IF 9.6 ) Pub Date : 2023-12-21 , DOI: 10.1021/acs.nanolett.3c03723
Bing Wang ₁ , Yi-Tong Xu ₁ , Tian-Yang Zhang ₁ , Hai-Yan Wang ₁ , Xian Zhang ₁ , Zeng-Qiang Wu ₂ , Wei-Wei Zhao ₁ , Hong-Yuan Chen ₁ , Jing-Juan Xu ₁

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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.

中文翻译：

由限制在纳米孔中的 CRISPR/Cas 驱动的超灵敏、高效的 microRNA 纳米传感器

这项工作提出了一种能够进行超灵敏 microRNA 检测的成簇规则间隔短回文重复序列 (CRISPR)/Cas-nanopipette 纳米电化学（Cas = CRISPR 相关蛋白）。将 CRISPR/Cas13a 纳米限制在纳米移液管内，可产生约 100% 的高催化效率。比本体电解质高 169 倍，有助于增强电化学响应。具有代表性的 microRNA-25 的 CRISPR/Cas13a 检测可实现低至 10 aM 的检测下限。该方法在单细胞和真实人血清检测中的实际应用得到了进一步证明。通过解决 microRNA-141 和 SARS-CoV-2 RNA 基因片段，验证了其普遍适用性。这项工作介绍了一种用于超灵敏纳米电化学和生物分析的新型 CRISPR/Cas 纳米技术。

更新日期：2023-12-21

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