Skip to content

Detecting RNA base methylations in single cells by in situ hybridization

Research output: Contribution to journalArticle

  • Rohan T. Ranasinghe
  • Martin R. Challand
  • Kristina A. Ganzinger
  • Benjamin W. Lewis
  • Charlotte Softley
  • Wolfgang H. Schmied
  • Mathew H. Horrocks
  • Nadia Shivji
  • Jason W. Chin
  • James Spencer
  • David Klenerman
Original languageEnglish
Article number655
Number of pages10
JournalNature Communications
Volume9
Issue number1
DOIs
DateAccepted/In press - 20 Dec 2017
DatePublished (current) - 13 Mar 2018

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 (m6 2A, 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.

Download statistics

No data available

Documents

Documents

  • Full-text PDF (final published version)

    Rights statement: This is the final published version of the article (version of record). It first appeared online via Nature at https://www.nature.com/articles/s41467-017-02714-7 . Please refer to any applicable terms of use of the publisher.

    Final published version, 1.44 MB, PDF document

    Licence: CC BY

  • Supplementary figure PDF

    Rights statement: This is the final published version of the article (version of record). It first appeared online via Nature at https://www.nature.com/articles/s41467-017-02714-7 . Please refer to any applicable terms of use of the publisher.

    Final published version, 6.44 MB, PDF document

    Licence: CC BY

DOI

View research connections

Related faculties, schools or groups