Synthesis of novel siRNAs having thymidine dimers consisting of a carbamate or a urea linkage at their 3′ overhang regions and their ability to suppress human RNase L protein expression

Yoshihito Ueno, Tomoharu Naito, Koshi Kawada, Aya Shibata, Hye Sook Kim, Yusuke Wataya, Yukio Kitade

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10 Citations (Scopus)

Abstract

In order to examine the effect of modifications at the 3′ overhang regions of short interfering RNAs (siRNAs) on their gene-silencing activities, we designed and synthesized novel siRNAs having thymidine dimers consisting of a carbamate or a urea linkage at their 3′ overhang regions. Suppression of human RNase L protein expression by these siRNAs was analyzed by immunoblot with RNase L-specific antibody. It was found that, at 24 h post-transfection, the modified siRNAs having the thymidine dimers with the carbamate and urea linkage suppress the protein expression 78 and 37 times more efficiently than that with the natural phosphodiester linkage, respectively. Furthermore, the siRNA containing the carbamate linkage was 37 times more resistant to nucleolytic degradation by snake venom phosphodiesterase than the siRNA consisting of the natural phosphodiester linkage. Thus, the RNA duplexes having the thymidine dimers with the carbamate or urea linkage at their 3′ overhang regions will be promising candidates for novel siRNA molecules to down-regulate protein expression.

Original languageEnglish
Pages (from-to)1168-1175
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume330
Issue number4
DOIs
Publication statusPublished - May 20 2005

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Keywords

  • Carbamate
  • RNAi
  • RNase L
  • Urea
  • siRNA

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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