Detecting RNA base methylations in single cells by in situ hybridization.
Schmied, Wolfgang H
MetadataShow full item record
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.. Nature communications, 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 are used to detect these modifications, reporting the average methylation state of ~104-107 cells, 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)
Embargo Lift Date
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