Vectors expressing efficient RNA decoys achieve the long-term suppression of specific microRNA activity in mammalian cells.
Whereas the strong and stable suppression of specific microRNA activity would be essential for the functional analysis of these molecules, and also for the development of therapeutic applications, effective inhibitory methods to achieve this have not yet been fully established. In our current study, we tested various RNA decoys which were designed to efficiently expose indigestible complementary RNAs to a specific miRNA molecule. These inhibitory RNAs were at the same time designed to be expressed in lentiviral vectors and to be transported into the cytoplasm after transcription by RNA polymerase III. We report the optimal conditions that we have established for the design of such RNA decoys (we term these molecules TuD RNAs; tough decoy RNAs). We finally demonstrate that TuD RNAs induce specific and strong biological effects and also show that TuD RNAs achieve the efficient and long-term-suppression of specific miRNAs for over 1 month in mammalian cells.
Haraguchi
Takeshi
T
Division of Host-Parasite Interaction, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
Ozaki
Yuka
Y
Iba
Hideo
H
D001665:Binding Sites; D002460:Cell Line; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D035683:MicroRNAs / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D022661:RNA, Untranslated / Q000096:biosynthesis* / Q000737:chemistry* / Q000378:metabolism; D014161:Transduction, Genetic
D035683:MicroRNAs; D022661:RNA, Untranslated