Small interfering RNAs containing full 2'-O-methylribonucleotide-modified sense strands display Argonaute2/eIF2C2-dependent activity.
RNA interference (RNAi) is a process by which short interfering RNAs (siRNAs) direct the degradation of complementary single-strand RNAs. In this study, we investigated the effects of full-strand phosphorothioate (PS) backbone and 2'-O-methyl (2'-OMe) sugar modifications on RNAi-mediated silencing. In contrast to previous reports, we have identified active siRNA duplexes containing full 2'-OMe-modified sense strands that display comparable activity to the unmodified analog of similar sequence. The structure of these modified siRNAs is the predominant determinant of their activity, with sequence and backbone composition being secondary. We further show, by using biotin-tagged siRNAs and affinity-tagged hAgo2/eIF2C2, that activity of siRNA duplexes containing full 2'-OMe substitutions in the sense strand is mediated by the RNA-induced silencing complex (RISC) and that strand-specific loading (or binding) to hAgo2 may be modulated through selective incorporation of these modifications.
Kraynack
Bryan A
BA
ISIS Pharmaceuticals, Carlsbad Research Center, 1896 Rutherford Road, Carlsbad, CA 92008, USA.
Baker
Brenda F
BF
D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D009711:Nucleotides / Q000032:analysis*; D010448:Peptide Initiation Factors / Q000737:chemistry* / Q000378:metabolism; D018075:RNA, Complementary; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
C466422:AGO2 protein, human; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D009711:Nucleotides; D010448:Peptide Initiation Factors; D018075:RNA, Complementary; D034741:RNA, Small Interfering