Enzymatic synthesis and RNA interference of nucleosides incorporating stable isotopes into a base moiety. Issue 20 (15th October 2015)

• Record Type: Journal Article
• Authors: Hatano, Akihiko; Shiraishi, Mitsuya; Terado, Nanae; Tanabe, Atsuhiro; Fukuda, Kenji
• Is Part Of: Bioorganic & medicinal chemistry. Volume 23:Issue 20(2015)
• Page Start: 6683
• Page End: 6688
• Publication Date: 2015-10-15
• Subjects: Thymidine phosphorylase -- Nucleoside synthesis -- Nucleosides incorporating stable isotopes -- 13C NMR -- RNA interference; Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; Biochemistry -- Periodicals; Chemistry, Clinical -- Periodicals; Chemistry, Organic -- Periodicals; Chimie bio-organique -- Périodiques; Chimie pharmaceutique -- Périodiques; 615.19
• DOI: 10.1016/j.bmc.2015.09.011
• Languages: English
• ISSNs: 0968-0896

Abstract

Graphical abstract: 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. 13 C NMR spectra showed spin–spin coupling between 13 C and 15 N 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.