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Highly Effective and Low-Cost MicroRNA Detection with CRISPR-Cas9
ACS Synthetic Biology ( IF 4.7 ) Pub Date : 2018-02-27 00:00:00 , DOI: 10.1021/acssynbio.7b00446
Xin-Yuan Qiu 1 , Lv-Yun Zhu 1 , Chu-Shu Zhu 1 , Jia-Xin Ma 1 , Tao Hou 2 , Xiao-Min Wu 1 , Si-Si Xie 1 , Lu Min 1 , De-An Tan 3 , Dong-Yi Zhang 1 , Lingyun Zhu 1
Affiliation  

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.

中文翻译:

使用CRISPR-Cas9进行高效且低成本的MicroRNA检测

据报道,MicroRNA与多种疾病有关,在诊断和治疗中具有潜在的应用。然而,鉴于目前的复杂性,高成本和低灵敏度,对miRNA的检测仍然是无法改进的。在这项研究中,我们试图建立一个新颖的平台,以低成本和高效率检测miRNA。该检测系统包含等温扩增,基于滚动环扩增,CRISPR-Cas9和辣根过氧化物酶技术的检测和报告过程。它仅具有单碱基特异性就可以检测样品中的痕量miRNA。而且,我们证明了该方案可有效检测临床血清样品中的靶标miRNA,并通过检测先前报道的生物标记物:循环let-7a,将非小细胞肺癌患者与健康志愿者区分开。作为第一个将CRISPR-Cas9用于miRNA检测的方法,该方法是一种有前途的方法,可用于多种疾病的筛查,诊断和预后。
更新日期:2018-02-27
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