Detecting RNA base methylations in single cells by in situ hybridization.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 29440632.
- Also identified by DOI 10.1038/s41467-017-02714-7 and PMC identifier 5811446.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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 ~10<sup>4</sup>-10<sup>7</sup> 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<sup>6</sup><sub>2</sub>A, m<sup>1</sup>G and m<sup>3</sup>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 multicolor fluorescence imaging.
Medical subject headings
- In Situ Hybridization, Fluorescence
- RNA
- RNA, Ribosomal
- Single-Cell Analysis