Arora Pearl, Zheng Haiyan, Munusamy Sathishkumar, Jahani Rana, Guan Xiyun
Department of Chemistry, Illinois Institute of Technology, Chicago, Illinois, USA.
Department of Chemistry, University of Missouri, Columbia, Missouri, USA.
Electrophoresis. 2024 Nov;45(21-22):2034-2044. doi: 10.1002/elps.202400134. Epub 2024 Aug 21.
MicroRNAs (miRNAs) play important roles in posttranscriptional gene regulation. Aberrations in the miRNA levels have been the cause behind various diseases, including periodontitis. Therefore, sensitive, specific, and accurate detection of disease-associated miRNAs is vital to early diagnosis and can facilitate inhibitor screening and drug design. In this study, we developed a label-free, real-time sensing method for the detection of miR31, which has been frequently linked to periodontitis, using an engineered protein nanopore and in the presence of a complementary ssDNA as a molecular probe. Our method is rapid and highly sensitive with nanomolar concentration of miR31 that could be determined in minutes. Furthermore, our sensor showed high selectivity toward the target miR31 sequence even in the presence of interfering nucleic acids. In addition, artificial saliva and human saliva samples were successfully analyzed. Our developed nanopore sensing platform could be used to detect other miRNAs and offers a potential application for the clinical diagnosis of disease biomarkers.
微小RNA（miRNA）在转录后基因调控中发挥着重要作用。miRNA水平的异常是包括牙周炎在内的各种疾病的病因。因此，灵敏、特异且准确地检测与疾病相关的miRNA对于早期诊断至关重要，并且有助于抑制剂筛选和药物设计。在本研究中，我们开发了一种无标记的实时传感方法，用于检测经常与牙周炎相关的miR31，该方法使用工程化蛋白质纳米孔，并在存在互补单链DNA作为分子探针的情况下进行。我们的方法快速且高度灵敏，能够在数分钟内测定纳摩尔浓度的miR31。此外，即使在存在干扰核酸的情况下，我们的传感器对目标miR31序列仍表现出高选择性。另外，成功分析了人工唾液和人类唾液样本。我们开发的纳米孔传感平台可用于检测其他miRNA，并为疾病生物标志物的临床诊断提供了潜在应用。
