Synthesis of RNA Crosslinking Oligonucleotides Modified with 2-Amino-7-Deaza-7-Propynyl-6-Vinylpurine

Curr Protoc Nucleic Acid Chem. 2019 Jun;77(1):e79. doi: 10.1002/cpnc.79. Epub 2019 Mar 12.

Abstract

This article describes procedures to synthesize 2'-OMe-RNA modified with cross-linkable 2-amino-7-deaza-7-propynyl-6-vinylpurine (ADpVP) and preparation of the RNA-crosslinking experiment in vitro. All synthesis steps yield the desired compound in moderate or high yield without expensive chemical reagents or specific devices. The crosslink-active form of modified RNA can also be purified by commonly used reversed-phase HPLC, can be stored at -80°C after lyophilization for a few days, and is ready to use for crosslinking experiments. This crosslink-active RNA can efficiently form covalent bonds with complementary RNA in a sequence-specific manner. © 2019 by John Wiley & Sons, Inc.

Keywords: 7-deaza-guanosine; RNA crosslink; nucleoside; oligonucleotide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Cross-Linking Reagents / chemistry
  • Oligonucleotides / chemical synthesis*
  • Oligonucleotides / chemistry
  • Purines / chemistry*
  • RNA / chemistry*
  • Vinyl Compounds / chemistry*

Substances

  • 2-amino-6-vinylpurine
  • Cross-Linking Reagents
  • Oligonucleotides
  • Purines
  • Vinyl Compounds
  • RNA