A novel synthesis and detection method for cap-associated adenosine modifications in mouse mRNA

Sci Rep. 2011:1:126. doi: 10.1038/srep00126. Epub 2011 Oct 24.

Abstract

A method is described for the detection of certain nucleotide modifications adjacent to the 5' 7-methyl guanosine cap of mRNAs from individual genes. The method quantitatively measures the relative abundance of 2'-O-methyl and N(6),2'-O-dimethyladenosine, two of the most common modifications. In order to identify and quantitatify the amounts of N(6),2'-O-dimethyladenosine, a novel method for the synthesis of modified adenosine phosphoramidites was developed. This method is a one step synthesis and the product can directly be used for the production of N(6),2'-O-dimethyladenosine containing RNA oligonucleotides. The nature of the cap-adjacent nucleotides were shown to be characteristic for mRNAs from individual genes transcribed in liver and testis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / chemistry
  • Animals
  • Base Sequence
  • Chromatography, Thin Layer
  • Liver / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Structure
  • RNA Cap Analogs / chemical synthesis*
  • RNA Cap Analogs / chemistry
  • RNA Cap Analogs / genetics
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Testis / metabolism

Substances

  • RNA Cap Analogs
  • RNA, Messenger
  • Adenosine