Direct microRNA Sequencing Using Nanopore-Induced Phase-Shift Sequencing

• Published in: iScience Vol. 23; no. 3; p. 100916
• Main Authors: Zhang, Jinyue, Yan, Shuanghong, Chang, Le, Guo, Weiming, Wang, Yuqin, Wang, Yu, Zhang, Panke, Chen, Hong-Yuan, Huang, Shuo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 27.03.2020; Elsevier
• Subjects: Analytical Chemistry; Biotechnology; Molecular Biology; Nanotechnology; Biotechnology; Analytical Chemistry; Molecular Biology; Nanotechnology
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2020.100916

MicroRNAs (miRNAs) are a class of short non-coding RNAs that function in RNA silencing and post-transcriptional gene regulation. However, direct characterization of miRNA is challenging due to its unique properties such as its low abundance, sequence similarities, and short length. Although urgently needed, single molecule sequencing of miRNA has never been demonstrated, to the best of our knowledge. Nanopore-induced phase-shift sequencing (NIPSS), which is a variant form of nanopore sequencing, could directly sequence any short analytes including miRNA. In practice, NIPSS clearly discriminates between different identities, isoforms, and epigenetic variants of model miRNA sequences. This work thus demonstrates direct sequencing of miRNA, which serves as a complement to existing miRNA sensing routines by the introduction of the single molecule resolution. Future engineering of this technique may assist miRNA-based early stage diagnosis or inspire novel cancer therapeutics. [Display omitted] •The first demonstration of single molecule miRNA sequencing•miRNA sequencing by NIPSS can directly identify epigenetic modifications•Enzymatic conjugation enables NIPSS sequencing of natural miRNAs Analytical Chemistry; Molecular Biology; Biotechnology; Nanotechnology