反式激活crRNA
清脆的
计算生物学
生物
核酸
条形码
核酸检测
遗传学
回文
计算机科学
基因
操作系统
作者
Miaojin Zhou,Chunhua Zhang,Miaomiao Chen,Zhiqing Hu,Menglin Li,Zhuo Li,Lingqian Wu,Desheng Liang
出处
期刊:MedComm
[Wiley]
日期:2023-07-02
卷期号:4 (4)
摘要
Abstract Clustered regularly interspaced short palindromic repeat (CRISPR)‐based biosensors have been developed to facilitate the rapid and sensitive detection of nucleic acids. However, most approaches using CRISPR‐based detection have disadvantages associated with the limitations of CRISPR RNA (crRNA), protospacer adjacent motif (PAM) or protospacer flanking sequence restriction, single channel detection, and difficulty in quantitative detection resulting in only some target sites being detected qualitatively. Here, we aimed to develop a barcode‐based Cas12a‐mediated DNA detection (BCDetection) strategy, which overcomes the aforementioned drawbacks and enables (1) detection with a universal PAM and crRNA without PAM or crRNA restriction, (2) simultaneous detection of multiple targets in a single reaction, and (3) quantitative detection, which can significantly distinguish copy number differences up to as low as a two‐fold limit. We could efficiently and simultaneously detect three β‐thalassemia mutations in a single reaction using BCDetection. Notably, samples from normal individuals, spinal muscular atrophy (SMA) carriers, and SMA patients were significantly and accurately distinguished using the quantitative detection ability of BCDetection, indicating its potential application in β‐thalassemia and SMA carrier screening. Therefore, our findings demonstrate that BCDetection provides a new platform for accurate and efficient quantitative detection using CRISPR/Cas12a, highlighting its bioanalytical applications.
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