RNA interference in mammalian cells by chemically-modified RNA

Dwaine A. Braasch, Susan Jensen, Yinghui Liu, Kiran Kaur, Khalil Arar, Michael A. White, David R. Corey

Research output: Contribution to journalArticle

437 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)7967-7975
Number of pages9
JournalBiochemistry
Volume42
Issue number26
DOIs
StatePublished - Jul 8 2003

ASJC Scopus subject areas

  • Biochemistry

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