Double-stranded regions are essential design components of potent inhibitors of RISC function.
While microRNAs (miRNAs) are recognized as playing a critical role in regulating eukaryotic gene expression, both the mechanism by which these small, noncoding RNAs function and the genes they target remain elusive. Previous studies have shown that short, single-stranded 2'-O-methyl-modified oligonucleotides that are complementary to mature microRNA sequences can interact with the miRNA-RISC nucleoprotein complex and weakly inhibit miRNA function. Here we report the identification of secondary structural elements that enhance the potency of these molecules. Incorporation of highly structured, double-stranded flanking regions around the reverse complement core significantly increases inhibitor function and allows for multi-miRNA inhibition at subnanomolar concentrations. The improved functionality of these double-stranded miRNA inhibitors may provide insights into the miRNA mechanism by suggesting the possible importance of such structures in or near endogenous miRNA target sites.
Vermeulen
Annaleen
A
Thermo Fisher Scientific, Dharmacon Products, Lafayette, Colorado 80026, USA. annaleen.vermeulen@thermofischer.com
Robertson
Barbara
B
Dalby
Andrew B
AB
Marshall
William S
WS
Karpilow
Jon
J
Leake
Devin
D
Khvorova
Anastasia
A
Baskerville
Scott
S
D002460:Cell Line; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D005821:Genetic Techniques; D006801:Humans; D035683:MicroRNAs / Q000037:antagonists & inhibitors*; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics; D012330:RNA, Double-Stranded / Q000737:chemistry*; D034743:RNA-Induced Silencing Complex / Q000037:antagonists & inhibitors*; D013329:Structure-Activity Relationship
D004791:Enzyme Inhibitors; D035683:MicroRNAs; D016372:RNA, Antisense; D012330:RNA, Double-Stranded; D034743:RNA-Induced Silencing Complex