期刊
ACS SYNTHETIC BIOLOGY
卷 7, 期 3, 页码 807-813出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.7b00446
关键词
miRNA detection; CRISPR-dCas9; rolling circle amplification; split reporter; biomarker
资金
- National Natural Science Foundation of China [31500686, 31100609]
- Hunan Provincial Natural Science Foundation [2017JJ3358]
- National University of Defense Technology project [JC14-02-09, ZK16-03-13]
MicroRNAs have been reported as related to multiple diseases and have potential applications in diagnosis and therapeutics. However, detection of rniRNAs remains improvable, given their complexity, high cost, and low sensitivity as of currently. In this study, we attempt to build a novel platform that detects miRNAs at low cost and high efficacy. This detection system contains isothermal amplification, detecting and reporting process based on rolling circle amplification, CRISPR-Cas9, and splitThorseradish peroxidase techniques. It is able to detect trace amount of miRNAs from samples with mere single-base specificity. Moreover, we demonstrated that such scheme can effectively detec target miRNAs in clinical serum samples and significantly distinguish patients of non-small cell lung cancer from healthy volunteers by detecting the previously reported biomarker: circulating let-7a. As the first to use CRISPR-Cas9 in miRNA detection, this method is a promising approach capable of being applied in screening, diagnosing, and prognosticating of multiple diseases.
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