Suppression of microRNAs by dual-targeting and clustered Tough Decoy inhibitors
Suppression of microRNAs by dual-targeting and clustered Tough Decoy inhibitors
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DOI：
10.4161/rna.23543
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发表时间：
2013-03-01
期刊：
影响因子：
4.1
通讯作者：
Mikkelsen, Jacob Giehm
中科院分区：
文献类型：
--
作者：
Hollensen, Anne Kruse;Bak, Rasmus O.;Mikkelsen, Jacob Giehm
MicroRNA (miRNA) 是普遍存在的基因表达调节因子，对几乎所有细胞过程都有贡献。管理 miRNA 活性的方法在各种实验和治疗应用中越来越受到关注。从基于质粒或病毒的载体表达的 DNA 编码的 miRNA 抑制剂提供持久的 miRNA 抑制和组织导向的微观管理选项。在本报告中，我们探索了利用短发夹状 RNA 同时抑制两个或多个 miRNA 的潜力。基于“Tough Decoy”（TuD）设计，我们创建了携带两个 miRNA 识别位点的双靶向发夹，并证明了单个 DNA 编码的抑制剂 RNA 对不同对不相关 miRNA 的有效共抑制。此外，通过表达携带簇状 TuD 发夹（总共具有多达 8 个 miRNA 识别位点）的 RNA 聚合酶 II 转录抑制剂，可以增强 miRNA 抑制作用。值得注意的是，通过表达对总共六个 miRNA 中的每一个具有单个识别位点的簇状 TuD 抑制剂，我们记录了由单个表达盒编码的抑制剂 RNA 分子对多个 miRNA 的强大并行抑制。这些发现揭示了基于 TuD 的 miRNA 抑制剂的新潜力，并为标准化 miRNA 家族或簇的同步抑制铺平了道路。
MicroRNAs (miRNAs) are ubiquitous regulators of gene expression that contribute to almost any cellular process. Methods for managing of miRNA activity are attracting increasing attention in relation to diverse experimental and therapeutic applications. DNA-encoded miRNA inhibitors expressed from plasmid or virus-based vectors provide persistent miRNA suppression and options of tissue-directed micromanaging. In this report, we explore the potential of exploiting short, hairpin-shaped RNAs for simultaneous suppression of two or more miRNAs. Based on the "Tough Decoy" (TuD) design, we create dual-targeting hairpins carrying two miRNA recognition sites and demonstrate potent co-suppression of different pairs of unrelated miRNAs by a single DNA-encoded inhibitor RNA. In addition, enhanced miRNA suppression is achieved by expression of RNA polymerase II-transcribed inhibitors carrying clustered TuD hairpins with up to a total of eight miRNA recognition sites. Notably, by expressing clustered TuD inhibitors harboring a single recognition site for each of a total of six miRNAs, we document robust parallel suppression of multiple miRNAs by inhibitor RNA molecules encoded by a single expression cassette. These findings unveil a new potential of TuD-based miRNA inhibitors and pave the way for standardizing synchronized suppression of families or clusters of miRNAs.