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ACS Synth Biol. 2018 Mar 16;7(3):807-813. doi: 10.1021/acssynbio.7b00446. Epub 2018 Mar 7.

Highly Effective and Low-Cost MicroRNA Detection with CRISPR-Cas9.

Author information

1
Department of Biology and Chemistry, College of Liberal Arts and Sciences , National University of Defense Technology , Changsha , Hunan 410073 , People's Republic of China.
2
Department of Oncology , The Second Xiangya Hospital of Central South University , Changsha , Hunan 410012 , People's Republic of China.
3
Department of Clinical Laboratory , Hospital of National University of Defense Technology , Changsha , Hunan 410073 , People's Republic of China.

Abstract

MicroRNAs have been reported as related to multiple diseases and have potential applications in diagnosis and therapeutics. However, detection of miRNAs 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 split-horseradish 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 detect 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.

KEYWORDS:

CRISPR-dCas9; biomarker; miRNA detection; rolling circle amplification; split reporter

PMID:
29486117
DOI:
10.1021/acssynbio.7b00446
[Indexed for MEDLINE]

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