Single-molecule RNA sequencing for simultaneous detection of m6A and 5mC.

Abstract

Epitranscriptomics is the study of RNA base modifications involving functionally relevant changes to the transcriptome. In recent years, epitranscriptomics has been an active area of research. However, a major issue has been the development of sequencing methods to map transcriptome-wide RNA base modifications. We have proposed a single-molecule quantum sequencer for mapping RNA base modifications in microRNAs (miRNAs), such as N6-methyladenosine (m6A) or 5-methylcytidine (5mC), which are related to cancer cell propagation and suppression. Here, we investigated 5mC and m6A in hsa-miR-200c-5p extracted from colorectal cancer cells and determined their methylation sites and rates; the data were comparable to those determined by mass spectrometry. Furthermore, we evaluated the methylation ratio of cytidine and adenosine at each site in the sequences and its relationship. These results suggest that the methylation ratio of cytidine and adenosine is facilitated by the presence of vicinal methylation. Our work provides a robust new tool for sequencing various types of RNA base modifications in their RNA context.

• Authors: Takahito Ohshiro, Masamitsu Konno, Ayumu Asai, Yuki Komoto, Akira Yamagata, Yuichiro Doki, Hidetoshi Eguchi, Ken Ofusa, Masateru Taniguchi, Hideshi Ishii
• Journal: Scientific reports (Sci Rep) Vol. 11 Issue 1 Pg. 19304 (09 29 2021) ISSN: 2045-2322 [Electronic] England
• PMID: 34588546 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2021. The Author(s).

Chemical References

• MIRN200 microRNA, human
• MicroRNAs
• Cytidine
• N-methyladenosine
• Adenosine
• 5-methylcytidine

Topics

• Adenosine (analogs & derivatives, isolation & purification, metabolism)
• Cell Line, Tumor
• Colorectal Neoplasms (genetics, pathology)
• Cytidine (analogs & derivatives, isolation & purification, metabolism)
• Epigenesis, Genetic
• Gene Expression Profiling (methods)
• Gene Expression Regulation, Neoplastic
• Humans
• Methylation
• MicroRNAs (chemistry, genetics, metabolism)
• Sequence Analysis, RNA (methods)
• Single Molecule Imaging (methods)