2. Academic Validation
  3. Detecting RNA base methylations in single cells by in situ hybridization

Detecting RNA base methylations in single cells by in situ hybridization

  • Nat Commun. 2018 Feb 13;9(1):655. doi: 10.1038/s41467-017-02714-7.
Rohan T Ranasinghe 1,  Martin R Challand 2 3,  Kristina A Ganzinger 4 5,  Benjamin W Lewis 4,  Charlotte Softley 4,  Wolfgang H Schmied 6,  Mathew H Horrocks 4,  Nadia Shivji 4,  Jason W Chin 4 6,  James Spencer 7,  David Klenerman 4
Affiliations

Affiliations

  • 1 Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. [email protected].
  • 2 School of Cellular and Molecular Medicine, University of Bristol, Bristol, BS8 1TD, UK. [email protected].
  • 3 School of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK. [email protected].
  • 4 Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
  • 5 Max-Planck-Institut für Biochemie (MPI for Biochemistry), 82152, Martinsried, Germany.
  • 6 Medical Research Council Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, CB2 0QH, UK.
  • 7 School of Cellular and Molecular Medicine, University of Bristol, Bristol, BS8 1TD, UK.
PMID: 29440632 DOI: 10.1038/s41467-017-02714-7
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.

Figures
Products