Detecting RNA base methylations in single cells by in situ hybridization.
Schmied, Wolfgang H
Springer Science and Business Media LLC
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Ranasinghe, R., Challand, M. R., Ganzinger, K. A., Lewis, B. W., Softley, C., Schmied, W. H., Horrocks, M., et al. (2018). Detecting RNA base methylations in single cells by in situ hybridization.. Nat Commun, 9 (1), 655. https://doi.org/10.1038/s41467-017-02714-7
Methylated bases in tRNA, rRNA and mRNA control a variety of cellular processes, including protein synthesis, antimicrobial resistance and gene expression. Currently, bulk methods that report the average methylation state of ~104-107 cells are used to detect these modifications, obscuring potentially important biological information. Here, we use in situ hybridization of Molecular Beacons for single-cell detection of three methylations (m62A, m1G and m3U) that destabilize Watson-Crick base pairs. Our method-methylation-sensitive RNA fluorescence in situ hybridization-detects single methylations of rRNA, quantifies antibiotic-resistant bacteria in mixtures of cells and simultaneously detects multiple methylations using multicolor fluorescence imaging.
Escherichia coli, Adenine, Guanine, Methyltransferases, Escherichia coli Proteins, RNA, RNA, Ribosomal, Uridine, Microscopy, Fluorescence, In Situ Hybridization, Fluorescence, Methylation, Single-Cell Analysis
European Commission (216027)
European Commission (115153)
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External DOI: https://doi.org/10.1038/s41467-017-02714-7
This record's URL: https://www.repository.cam.ac.uk/handle/1810/273301
Attribution 4.0 International, Attribution 4.0 International, Attribution 4.0 International, Attribution 4.0 International