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

Alison Galloway, Abdelmadjid Atrih, Renata Grzela, Edward Darzynkiewicz, Michael A. J. Ferguson (Lead / Corresponding author), Victoria H. Cowling (Lead / Corresponding author)

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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.

Original languageEnglish
Article number190306
Number of pages13
JournalOpen Biology
Volume10
Issue number2
DOIs
Publication statusPublished - 26 Feb 2020

Keywords

  • RNA processing
  • mass spectrometry
  • ribose O-2 methylation
  • RNA cap
  • 7-methylguanosine
  • RNA methylation
  • Mass spectrometry
  • Ribose O-2 methylation

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