Vectors expressing efficient RNA decoys achieve the long-term suppression of specific microRNA activity in mammalian cells
Open Access
- 17 February 2009
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 37 (6), e43
- https://doi.org/10.1093/nar/gkp040
Abstract
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.Keywords
This publication has 28 references indexed in Scilit:
- miR-21 Gene Expression Triggered by AP-1 Is Sustained through a Double-Negative Feedback MechanismJournal of Molecular Biology, 2008
- MicroRNA genes are transcribed by RNA polymerase IIThe EMBO Journal, 2004
- Structural requirements for pre-microRNA binding and nuclear export by Exportin 5Nucleic Acids Research, 2004
- Cre-lox-regulated conditional RNA interference from transgenesProceedings of the National Academy of Sciences, 2004
- CRE recombinase-inducible RNA interference mediated by lentiviral vectorsProceedings of the National Academy of Sciences of the United States of America, 2004
- Sequence-Specific Inhibition of Small RNA FunctionPLoS Biology, 2004
- A high-throughput method to monitor the expression of microRNA precursorsNucleic Acids Research, 2004
- Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencingRNA, 2004
- Refractory nature of normal human diploid fibroblasts with respect to oncogene-mediated transformationProceedings of the National Academy of Sciences, 2003
- A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applicationsNature Biotechnology, 2002