Enzymatic synthesis and RNA interference of nucleosides incorporating stable isotopes into a base moiety

Akihiko Hatano, Mitsuya Shiraishi, Nanae Terado, Atsuhiro Tanabe, Kenji Fukuda

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Thymidine phosphorylase was used to catalyze the conversion of thymidine (or methyluridine) and uracil incorporating stable isotopes to deoxyuridine (or uridine) with the uracil base incorporating the stable isotope. These base-exchange reactions proceeded with high conversion rates (75-96%), and the isolated yields were also good (64-87%). The masses of all synthetic compounds incorporating stable isotopes were identical to the theoretical molecular weights via EIMS. 13C NMR spectra showed spin-spin coupling between 13C and 15N in the synthetic compounds, and the signals were split, further proving incorporation of the isotopes into the compounds. The RNA interference effects of this siRNA with uridine incorporating stable isotopes were also investigated. A 25mer siRNA had a strong knockdown effect on the MARCKS protein. The insertion position and number of uridine moieties incorporating stable isotopes introduced into the siRNA had no influence on the silencing of the target protein. This incorporation of stable isotopes into RNA and DNA has the potential to function as a chemically benign tracer in cells.

Original languageEnglish
Pages (from-to)6683-6688
Number of pages6
JournalBioorganic and Medicinal Chemistry
Volume23
Issue number20
DOIs
Publication statusPublished - 2015 Oct 15

Keywords

  • C NMR
  • Nucleoside synthesis
  • Nucleosides incorporating stable isotopes
  • RNA interference
  • Thymidine phosphorylase

ASJC Scopus subject areas

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

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