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

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.

Towards the Discovery of Oligonucleotide Cross-Linking Agents

“…To overcome these limitations several derivatizations were carried out over vinylpurines with slightly better cross-linking properties. [70,72,73] Though some improvements have been made over the period, they are not suitable for the development of an antisense drug. [71] Modifications should be made to achieve irreversible covalent bond.…”

“…Previous attempts failed to provide a suitable leaving group for a nucleophilic attack to produce a permanent covalent bond. [59,[70][71][72][73] Absence of a leaving group is one of the main drawbacks of cross-linking agents in the past.…”