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Detecting RNA base methylations in single cells by in situ hybridization.

ABSTRACT: 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.

SUBMITTER: Ranasinghe RT

PROVIDER: S-EPMC5811446 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature

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Detecting RNA base methylations in single cells by in situ hybridization.

Ranasinghe Rohan T RT Challand Martin R MR Ganzinger Kristina A KA Lewis Benjamin W BW Softley Charlotte C Schmied Wolfgang H WH Horrocks Mathew H MH Shivji Nadia N Chin Jason W JW Spencer James J Klenerman David D

Nature communications 20180213 1

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, m1</ ...[more]

PMID: 29440632

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