Vectors expressing efficient RNA decoys achieve the long-term suppression of specific microRNA activity in mammalian cells.

Vectors expressing efficient RNA decoys achieve the long-term suppression of specific microRNA activity in mammalian cells.
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DOI:
10.1093/nar/gkp040
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发表时间:
2009-04
影响因子:
14.9
通讯作者:
Iba H
Iba H
中科院分区:
生物学2区
文献类型:
--
作者:
Haraguchi T;Ozaki Y;Iba H

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尽管对特定microRNA活性的强而稳定的抑制对于这些分子的功能分析以及治疗应用的发展至关重要,但实现这一目标的有效抑制方法尚未完全建立。在我们目前的研究中,我们测试了各种RNA诱饵,这些诱饵旨在有效地将不可消化的互补RNA暴露于特定的miRNA分子。这些抑制RNA同时被设计成在慢病毒载体上表达,并通过RNA聚合酶III转录后转运到细胞质中。我们报告了我们为设计这种RNA诱饵(我们将这些分子称为TuD RNA;坚韧的诱饵RNA)建立的最佳条件。我们最终证明了TuD rna诱导特异性和强大的生物学效应,并表明TuD rna在哺乳动物细胞中实现了对特异性mirna的有效和长期抑制,持续时间超过1个月。
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.
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