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Detecting RNA base methylations in single cells by in situ hybridization

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

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Zitation

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.


Zitierlink: https://hdl.handle.net/21.11116/0000-0000-F6F2-9
Zusammenfassung
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.