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CRISPR/Cas12a介导的脂质体扩增策略用于核酸的光电化学检测

CRISPR/Cas12a-mediated liposome-amplified strategy for the photoelectrochemical detection of nucleic acid.

作者信息

Gong Hexiang, Wu Yulin, Zeng Ruijin, Zeng Yongyi, Liu Xiaolong, Tang Dianping

机构信息

Key Laboratory for Analytical Science of Food Safety and Biology (MOE & Fujian Province), State Key Laboratory of Photocatalysis on Energy and Environment, Department of Chemistry, Fuzhou University, Fuzhou 350108, China.

The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou 350025, China.

出版信息

Chem Commun (Camb). 2021 Sep 6;57(71):8977-8980. doi: 10.1039/d1cc03743a.

Abstract

This study reports a photoelectrochemical biosensor for dopamine-loaded liposome-encoded magnetic beads cleaved by clustered regularly interspaced short palindromic repeats (CRISPR)/Cas 12a system for the quantification of human papilloma virus (HPV)-related DNA using neodymium-doped BiOBr nanosheets (Nd-BiOBr) as a photoactive matrix. Magnetic beads and dopamine-loaded liposomes are covalently attached to the both ends of ssDNA to construct dumbbell-shaped dopamine-loaded liposome-encoded magnetic bead (DLL-MB) probes. When the guide RNA binds to the target HPV-16, the ssDNA will be cleaved by Cas12a, thereby degrading the double dumbbell probes. After magnetic separation, the dissolved DLLs are treated with Triton X-100 to release the dopamine (as an electron donor), which was then detected by an amplified photocurrent using the Nd-BiOBr-based photoelectrode.

摘要

本研究报道了一种用于载多巴胺脂质体编码磁珠的光电化学生物传感器,该磁珠由成簇规律间隔短回文重复序列(CRISPR)/Cas 12a系统切割,以掺钕溴氧化铋纳米片(Nd-BiOBr)作为光活性基质来定量人乳头瘤病毒(HPV)相关DNA。磁珠和载多巴胺脂质体共价连接到单链DNA的两端,构建哑铃状载多巴胺脂质体编码磁珠(DLL-MB)探针。当向导RNA与目标HPV-16结合时,单链DNA将被Cas12a切割,从而降解双哑铃探针。磁分离后,用曲拉通X-100处理溶解的DLL以释放多巴胺(作为电子供体),然后使用基于Nd-BiOBr的光电极通过放大的光电流对其进行检测。

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