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Journal Article

Detecting RNA base methylations in single cells by in situ hybridization


Ganzinger,  Kristina A.
Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society;

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Ranasinghe, R. T., Challand, M. R., Ganzinger, K. A., Lewis, B. W., Softley, C., Schmied, W. H., et al. (2018). Detecting RNA base methylations in single cells by in situ hybridization. Nature Communications, 9: 655. doi:10.1038/s41467-017-02714-7.

Cite as: https://hdl.handle.net/21.11116/0000-0000-F6F2-9
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 -10(4)-10(7) 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 (m(2)(6)A, m(1)G and m(3)U) 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 multi-color fluorescence imaging.