CAP-MAP: cap analysis protocol with minimal analyte processing, a rapid and sensitive approach to analysing mRNA cap structures

Open Biol. 2020 Feb;10(2):190306. doi: 10.1098/rsob.190306. Epub 2020 Feb 26.

Abstract

Eukaryotic messenger RNA (mRNA) is modified by the addition of an inverted guanosine cap to the 5' triphosphate. The cap guanosine and initial transcribed nucleotides are further methylated by a series of cap methyltransferases to generate the mature cap structures which protect RNA from degradation and recruit proteins involved in RNA processing and translation. Research demonstrating that the cap methyltransferases are regulated has generated interest in determining the methylation status of the mRNA cap structures present in cells. Here, we present CAP-MAP: cap analysis protocol with minimal analyte processing, a rapid and sensitive method for detecting cap structures present in mRNA isolated from tissues or cultured cells.

Keywords: 7-methylguanosine; RNA cap; RNA methylation; RNA processing; mass spectrometry; ribose O-2 methylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Chromatography, Liquid
  • Guanosine / metabolism
  • Liver / chemistry
  • Liver / cytology*
  • Mass Spectrometry
  • Methyltransferases / metabolism*
  • Mice
  • Molecular Structure
  • RNA Caps / analysis*
  • RNA Caps / chemistry

Substances

  • RNA Caps
  • Guanosine
  • Methyltransferases