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