Qiu Xin-Yuan, Zhu Lv-Yun, Zhu Chu-Shu, Ma Jia-Xin, Hou Tao, Wu Xiao-Min, Xie Si-Si, Min Lu, Tan De-An, Zhang Dong-Yi, Zhu Lingyun
Department of Biology and Chemistry, College of Liberal Arts and Sciences , National University of Defense Technology , Changsha , Hunan 410073 , People's Republic of China.
Department of Oncology , The Second Xiangya Hospital of Central South University , Changsha , Hunan 410012 , People's Republic of China.
ACS Synth Biol. 2018 Mar 16;7(3):807-813. doi: 10.1021/acssynbio.7b00446. Epub 2018 Mar 7.
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.
据报道，微小RNA与多种疾病相关，在诊断和治疗方面具有潜在应用价值。然而，鉴于微小RNA目前的复杂性、高成本和低灵敏度，其检测仍有待改进。在本研究中，我们试图构建一个低成本、高效能检测微小RNA的新型平台。该检测系统包含基于滚环扩增、CRISPR-Cas9和分裂辣根过氧化物酶技术的等温扩增、检测和报告过程。它能够仅以单碱基特异性从样品中检测痕量微小RNA。此外，我们证明了该方案能够有效检测临床血清样品中的靶标微小RNA，并通过检测先前报道的生物标志物：循环中的let-7a，显著区分非小细胞肺癌患者与健康志愿者。作为首个将CRISPR-Cas9用于微小RNA检测的方法，该方法是一种有前景的方法，能够应用于多种疾病的筛查、诊断和预后评估。
