Long-read single-molecule RNA structure sequencing using nanopore.

Autor:	Bizuayehu TT; Computational Biology Unit, Department of Informatics, University of Bergen, Norway.; Sars International Center for Marine Molecular Biology, University of Bergen, Norway., Labun K; Computational Biology Unit, Department of Informatics, University of Bergen, Norway., Jakubec M; Department of Chemistry, University of Tromsø, Norway., Jefimov K; Computational Biology Unit, Department of Informatics, University of Bergen, Norway., Niazi AM; Computational Biology Unit, Department of Informatics, University of Bergen, Norway., Valen E; Computational Biology Unit, Department of Informatics, University of Bergen, Norway.; Sars International Center for Marine Molecular Biology, University of Bergen, Norway.
Jazyk:	angličtina
Zdroj:	Nucleic acids research [Nucleic Acids Res] 2022 Nov 11; Vol. 50 (20), pp. e120.
DOI:	10.1093/nar/gkac775
Abstrakt:	RNA molecules can form secondary and tertiary structures that can regulate their localization and function. Using enzymatic or chemical probing together with high-throughput sequencing, secondary structure can be mapped across the entire transcriptome. However, a limiting factor is that only population averages can be obtained since each read is an independent measurement. Although long-read sequencing has recently been used to determine RNA structure, these methods still used aggregate signals across the strands to detect structure. Averaging across the population also means that only limited information about structural heterogeneity across molecules or dependencies within each molecule can be obtained. Here, we present Single-Molecule Structure sequencing (SMS-seq) that combines structural probing with native RNA sequencing to provide non-amplified, structural profiles of individual molecules with novel analysis methods. Our new approach using mutual information enabled single molecule structural interrogation. Each RNA is probed at numerous bases enabling the discovery of dependencies and heterogeneity of structural features. We also show that SMS-seq can capture tertiary interactions, dynamics of riboswitch ligand binding, and mRNA structural features. (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)
Databáze:	MEDLINE
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