RNA interference in mammalian cells by chemically-modified RNA
RNA interference in mammalian cells by chemically-modified RNA
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DOI：
10.1021/bi0343774
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发表时间：
2003-07-08
期刊：
影响因子：
2.9
通讯作者：
Corey, DR
中科院分区：
文献类型：
--
作者：
Braasch, DA;Jensen, S;Corey, DR
RNA干扰（RNAi）被证明是控制哺乳动物细胞中基因表达的一种强大而通用的技术。然而，为了充分实现其在体内的潜力，可能需要引入化学修饰以优化效力、稳定性和药代动力学性质。在这里，我们测试了化学修饰对RNA稳定性和基因表达抑制的影响。我们发现，含有磷酸二酯或不同数量的硫代磷酸酯键的RNA双链体在血清中长时间孵育期间非常稳定。用含有硫代磷酸酯键的RNA双链体处理细胞导致基因表达的选择性抑制。RNAi还耐受2 '-脱氧-2'-氟尿苷或锁核酸（LNA）核苷酸的引入。LNA核苷酸的引入还显著增加了修饰的RNA双链体的热稳定性，而不损害RNAi的效率。这些结果表明，通过RNAi抑制基因表达与对双链体的广谱化学修饰是相容的，从而为探测RNAi机制和改善体内RNA干扰提供了广泛的有用选择。
RNA interference (RNAi) is proving to be a robust and versatile technique for controlling gene expression in mammalian cells. To fully realize its potential in vivo, however, it may be necessary to introduce chemical modifications to optimize potency, stability, and pharmacokinetic properties. Here, we test the effects of chemical modifications on RNA stability and inhibition of gene expression. We find that RNA duplexes containing either phosphodiester or varying numbers of phosphorothioate linkages are remarkably stable during prolonged incubations in serum. Treatment of cells with RNA duplexes containing phosphorothioate linkages leads to selective inhibition of gene expression. RNAi also tolerates the introduction of 2'-deoxy-2'-fluorouridine or locked nucleic acid (LNA) nucleotides. Introduction of LNA nucleotides also substantially increases the thermal stability of modified RNA duplexes without compromising the efficiency of RNAi. These results suggest that inhibition of gene expression by RNAi is compatible with a broad spectrum of chemical modifications to the duplex, affording a wide range of useful options for probing the mechanism of RNAi and for improving RNA interference in vivo.