Chemo-biological mRNA imaging with single nucleotide specificity

Chem Commun (Camb). 2019 Dec 5;55(98):14817-14820. doi: 10.1039/c9cc06989e.

Abstract

Unambiguous imaging of C → U edited mRNA calls for a method that distinguishes a locally high concentration of unbound probe or single nucleotide mismatched target from a locally low concentration of matched mRNA target. To address this issue, we combine FIT probes as a "chemical" detection system with the "biological" MS2 technique. Ratio measurements provide a convenient parameter to discriminate the edited from the unedited state of mRNA encoding for GlyR α2 in HEK cells.

MeSH terms

  • Benzothiazoles / chemistry
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism
  • DNA Probes / chemistry
  • DNA Probes / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Levivirus / metabolism
  • Microscopy, Fluorescence
  • Quinolines / chemistry
  • RNA, Messenger / metabolism*
  • Receptors, Glycine / genetics

Substances

  • Benzothiazoles
  • Capsid Proteins
  • DNA Probes
  • Quinolines
  • RNA, Messenger
  • Receptors, Glycine
  • enhanced green fluorescent protein
  • thiazole orange
  • Green Fluorescent Proteins