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

Fingerprint

Nucleosides
Isotopes
RNA
Uridine
Small Interfering RNA
Uracil
Thymidine Phosphorylase
Deoxyuridine
Thymidine
Molecular weight
Nuclear magnetic resonance
DNA
Proteins

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

Cite this

Enzymatic synthesis and RNA interference of nucleosides incorporating stable isotopes into a base moiety. / Hatano, Akihiko; Shiraishi, Mitsuya; Terado, Nanae; Tanabe, Atsuhiro; Fukuda, Kenji.

In: Bioorganic and Medicinal Chemistry, Vol. 23, No. 20, 15.10.2015, p. 6683-6688.

Research output: Contribution to journalArticle

Hatano, Akihiko ; Shiraishi, Mitsuya ; Terado, Nanae ; Tanabe, Atsuhiro ; Fukuda, Kenji. / Enzymatic synthesis and RNA interference of nucleosides incorporating stable isotopes into a base moiety. In: Bioorganic and Medicinal Chemistry. 2015 ; Vol. 23, No. 20. pp. 6683-6688.
@article{55d8fc3ac612420fae91d8465e5d857f,
title = "Enzymatic synthesis and RNA interference of nucleosides incorporating stable isotopes into a base moiety",
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.",
keywords = "C NMR, Nucleoside synthesis, Nucleosides incorporating stable isotopes, RNA interference, Thymidine phosphorylase",
author = "Akihiko Hatano and Mitsuya Shiraishi and Nanae Terado and Atsuhiro Tanabe and Kenji Fukuda",
year = "2015",
month = "10",
day = "15",
doi = "10.1016/j.bmc.2015.09.011",
language = "English",
volume = "23",
pages = "6683--6688",
journal = "Bioorganic and Medicinal Chemistry",
issn = "0968-0896",
publisher = "Elsevier Limited",
number = "20",

}

TY - JOUR

T1 - Enzymatic synthesis and RNA interference of nucleosides incorporating stable isotopes into a base moiety

AU - Hatano, Akihiko

AU - Shiraishi, Mitsuya

AU - Terado, Nanae

AU - Tanabe, Atsuhiro

AU - Fukuda, Kenji

PY - 2015/10/15

Y1 - 2015/10/15

N2 - 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.

AB - 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.

KW - C NMR

KW - Nucleoside synthesis

KW - Nucleosides incorporating stable isotopes

KW - RNA interference

KW - Thymidine phosphorylase

UR - http://www.scopus.com/inward/record.url?scp=84943518042&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84943518042&partnerID=8YFLogxK

U2 - 10.1016/j.bmc.2015.09.011

DO - 10.1016/j.bmc.2015.09.011

M3 - Article

C2 - 26404411

AN - SCOPUS:84943518042

VL - 23

SP - 6683

EP - 6688

JO - Bioorganic and Medicinal Chemistry

JF - Bioorganic and Medicinal Chemistry

SN - 0968-0896

IS - 20

ER -