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  • Review Article
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siRNAs: applications in functional genomics and potential as therapeutics

Nature Reviews Drug Discovery volume 3, pages 318–329 (2004)Cite this article

Key Points

  • RNA interference (RNAi) is a post-transcriptional gene-silencing mechanism that utilizes short interfering RNAs (siRNAs) as effector molecules to guide target mRNA cleavage.

  • siRNA is a new class of nucleic-acid-based gene-silencing molecule that could be more potent than ribozymes and antisense oligonucleotides in silencing gene expression because it uses cellular machinery.

  • RNAi can be applied in mammalian cells through the application of a vector that transcribes a hairpin RNA that is processed into siRNAs in the cell, or by directly transfecting siRNA into cells.

  • The robustness of the RNAi approach has motivated numerous groups to conduct near-genome-wide screens in mammalian cells to identify and validate potential drug targets.

  • RNAi has become an alternative mode for studying gene function in mouse by either the creation of transgenic mice that express short hairpin RNAs or by local or systemic introduction of siRNA by various injection methods.

  • siRNAs have the potential for inducing non-specific and sequence-specific off-target effects if they are not properly selected and applied. New findings in the study of the mechanisms of RNAi have suggested rules for the selection of potent siRNA sequences.

  • Like ribozymes and ODNs, siRNAs are being developed for therapeutic applications. Along this line, numerous groups have identified chemical modifications that increase the stability of siRNAs, while maintaining a high gene-silencing efficiency.

Abstract

Molecules that can specifically silence gene expression are powerful research tools. Much effort has been put into the development of such molecules and has resulted in the creation of different classes of potential therapeutic agents. Small interfering RNA (siRNA) is one of the latest additions to the repertoire of sequence-specific gene-silencing agents. The robustness of this approach has motivated numerous biotechnology organizations and academic institutions to develop siRNA libraries for high-throughput genome-wide screening in mammalian cells. This article first overviews current nucleic-acid-based approaches for gene silencing, and then focuses on the application of siRNAs in particular in functional genomics and as potential therapeutics.

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Figure 1: Mechanisms of nucleic-acid-based approaches for gene silencing: RNA silencing.
Figure 2: Mechanisms of nucleic-acid-based approaches for gene silencing: antisense compounds.
Figure 3: Mechanisms of nucleic-acid-based approaches for gene silencing: ribozymes.
Figure 4: Features of efficient and specific siRNAs.
Figure 5: In vivo mammalian gene silencing.

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Acknowledgements

We would like to thank F. Eckstein and M. Manoharan for their thoughtful comments on the manuscript. We would also like to thank A. Patkaniowska for the Table of siRNA chemical modifications, and the members of the Tuschl laboratory for critical comments.

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  1. Laboratory of RNA Molecular Biology, Rockefeller University, 1230 York Avenue, Box 186, New York, 10021, New York, USA

    Yair Dorsett & Thomas Tuschl

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  1. Yair Dorsett
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Correspondence to Thomas Tuschl.

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Competing interests

T.T. is a co-founder of Alnylam Pharmaceuticals, which develops siRNA-based therapeutics.

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FURTHER INFORMATION

RNA interference web focus

Glossary

INTERFERON RESPONSE

A cellular response to dsRNA longer than 30 base pairs that results in global post-transcriptional gene silencing.

CHROMATIN

Complex of DNA, histones and non-histone proteins from which eukaryotic chromosomes are formed.

DOMINANT DISEASE

A disease caused by a dominant genetic mutation.

SATURATING GENETIC SCREEN

A screen of sufficient scale to identify all possible target genes.

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Dorsett, Y., Tuschl, T. siRNAs: applications in functional genomics and potential as therapeutics. Nat Rev Drug Discov 3, 318–329 (2004). https://doi.org/10.1038/nrd1345

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