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RNA interference in mammalian cells by chemically-modified RNA

机译：化学修饰的RNA干扰哺乳动物细胞中的RNA

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摘要

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. [References: 41]
机译：RNA干扰（RNAi）被证明是一种控制哺乳动物细胞中基因表达的强大而通用的技术。但是，要完全实现其在体内的潜力，可能有必要引入化学修饰以优化药效，稳定性和药代动力学特性。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，在血清中长时间孵育过程中，含有磷酸二酯或不同数量的硫代磷酸酯键的RNA双链体非常稳定。用含有硫代磷酸酯键的RNA双链体处理细胞会导致基因表达的选择性抑制。 RNAi还耐受2'-脱氧-2'-氟尿苷或锁定核酸（LNA）核苷酸的引入。 LNA核苷酸的引入也大大提高了修饰的RNA双链体的热稳定性，而不会影响RNAi的效率。这些结果表明，RNAi对基因表达的抑制与双链体的广泛化学修饰相容，为探索RNAi的机制和改善体内RNA干扰提供了广泛的有用选择。 [参考：41]

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