Synthesis of nuclease-resistant siRNAs possessing benzene-phosphate backbones in their 3'-overhang regions.
We describe the synthesis and silencing activities of siRNA possessing N(1)-[3,5-bis(hydroxymethyl)phenyl]thymine (b(t)) in their 3'-overhang regions. We found that an siRNA possessing b(t) in the 3'-overhang region was more effective than an siRNA with natural nucleosides and the siRNA possessing 1,3-bis(hydroxymethyl)benzene (b) without a nucleobase at the 3'-overhang region in in vitro experiment using HeLa cells system. Furthermore, the RNA possessing b(t) at its 3'-end was more resistant to nucleolytic hydrolysis by snake venom phosphodiesterase (a 3'-exonuclease) than the RNA possessing the natural nucleoside 2'-deoxythymidine at the 3'-end. Thus, the compound b(t) will be a novel 3'-overhang moiety that can enhance the silencing activity and nuclease-resistant property of siRNAs.
Ueno
Yoshihito
Y
Department of Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan. uenoy@gifu-u.ac.jp
Inoue
Takumi
T
Yoshida
Mahito
M
Yoshikawa
Kayo
K
Shibata
Aya
A
Kitamura
Yoshiaki
Y
Kitade
Yukio
Y
D001483:Base Sequence; D001555:Benzene Derivatives / Q000138:chemical synthesis* / Q000737:chemistry; D004722:Endoribonucleases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D006367:HeLa Cells; D006801:Humans; D009691:Nucleic Acid Denaturation; D010755:Organophosphates / Q000138:chemical synthesis* / Q000737:chemistry; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry; D034741:RNA, Small Interfering / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D012910:Snake Venoms / Q000201:enzymology; D013329:Structure-Activity Relationship; D013936:Thymidine / Q000737:chemistry*; D014407:Tumor Cells, Cultured; D014508:Urea / Q000737:chemistry*
D001555:Benzene Derivatives; D010755:Organophosphates; D034741:RNA, Small Interfering; D012910:Snake Venoms; D014508:Urea; D004722:Endoribonucleases; D010727:Phosphoric Diester Hydrolases; D013936:Thymidine