小RNA
化学
清脆的
核酸
生物传感器
计算生物学
反式激活crRNA
DNA
多重位移放大
乳腺癌
锁核酸
癌细胞
分子生物学
癌症
聚合酶链反应
生物
DNA提取
Cas9
基因
生物化学
遗传学
作者
Jiawei Peng,Ting Liu,Le Luo Guan,Ziyue Xu,Tao Xiong,Yu Zhang,Junlong Song,Xuexia Liu,Yong Yang,Xian Hao
出处
期刊:Talanta
[Elsevier]
日期:2024-06-01
卷期号:273: 125938-125938
标识
DOI:10.1016/j.talanta.2024.125938
摘要
The expression levels of microRNA (miRNA) vary significantly in correlation with the occurrence and progression of cancer, making them valuable biomarkers for cancer diagnosis. However, their quantitative detection faces challenges due to the high sequence homology, low abundance and small size. In this work, we established a strand displacement amplification (SDA) approach based on miRNA-triggered structural "Lock" nucleic acid ("Lock" DNA), coupled with the CRISPR/Cas12a system, for detecting miRNA-21 in breast cancer cells. The "Lock" DNA freed the CRISPR-derived RNA (crRNA) from the dependence on the target sequence and greatly facilitated the extended detection of different miRNAs. Moreover, the CRISPR/Cas12a system provided excellent amplification ability and specificity. The designed biosensor achieved high sensitivity detection of miRNA-21 with a limit of detection (LOD) of 28.8 aM. In particular, the biosensor could distinguish breast cancer cells from other cancer cells through intracellular imaging. With its straightforward sequence design and ease of use, the Lock-Cas12a biosensor offers significant advantages for cell imaging and early clinical diagnosis.
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