Synthesis of native-like crosslinked duplex RNA and study of its properties

Kazumitsu Onizuka, Madoka E. Hazemi, Justin M. Thomas, Leanna R. Monteleone, Ken Yamada, Shuhei Imoto, Peter A. Beal, Fumi Nagatsugi

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

A variety of enzymes have been found to interact with double-stranded RNA (dsRNA) in order to carry out its functions. We have endeavored to prepare the covalently crosslinked native-like duplex RNA, which could be useful for biochemical studies and RNA nanotechnology. In this study, the interstrand covalently linked duplex RNA was formed by a crosslinking reaction between vinylpurine (VP) and the target cytosine or uracil in RNA. We measured melting temperatures and CD spectra to identify the properties of the VP crosslinked duplex RNA. The crosslinking formation increased the thermodynamic stability without disturbing the natural conformation of dsRNA. In addition, a competitive binding experiment with the duplex RNA binding enzyme, ADAR2, showed the crosslinked dsRNA bound the protein with nearly the same binding affinity as the natural dsRNA, confirming that it has finely preserved the natural traits of duplex RNA.

Original languageEnglish (US)
Pages (from-to)2191-2199
Number of pages9
JournalBioorganic and Medicinal Chemistry
Volume25
Issue number7
DOIs
StatePublished - Apr 1 2017

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Keywords

  • 6-Vinylpurine
  • ADAR
  • Crosslinking reaction
  • Native-like
  • RNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Onizuka, K., Hazemi, M. E., Thomas, J. M., Monteleone, L. R., Yamada, K., Imoto, S., Beal, P. A., & Nagatsugi, F. (2017). Synthesis of native-like crosslinked duplex RNA and study of its properties. Bioorganic and Medicinal Chemistry, 25(7), 2191-2199. https://doi.org/10.1016/j.bmc.2017.02.034