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Specific and potent RNAi in the nucleus of human cells

Nature Structural & Molecular Biology volume 12, pages 133–137 (2005)Cite this article

Abstract

RNA interference (RNAi) has become a research tool to control gene expression in various organisms and holds potential as a new therapeutic strategy. The mechanism of small interfering RNA (siRNA)-mediated RNAi involves target mRNA cleavage and destruction in the cytoplasm. We investigated siRNA-mediated induction of RNAi in the nucleus of human cells. Notably, we observed highly efficient knockdown of small nuclear RNA 7SK by siRNA. siRNA- and microRNA-programmed RNA-induced silencing complexes (RISCs) were present in both cytoplasmic and nuclear compartments and specifically cleaved their perfectly matched target RNA with markedly high efficiencies. Our results provide the first evidence that human RISCs programmed with siRNA are present in the nucleus and can knock down target RNA levels. These studies reveal new roles for the RNAi machinery in modulating post-transcriptional gene expression in the nucleus.

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Figure 1: siRNA-targeted knockdown of 7SK snRNA.
Figure 2: siRNA-mediated knockdown of U6 snRNA.
Figure 3: Target RNA cleavage by nuclear and cytoplasmic RISC* in vitro.
Figure 4: RISC* components are present in nuclear and cytoplasmic compartments.

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Acknowledgements

We thank members of the Rana lab, C. Mello and P. Zamore for helpful discussions, and T.J. Richman for editorial assistance. We also thank T. Tuschl and G.J. Hannon for sharing valuable reagents. This work was supported by a grant from the US National Institutes of Health (AI43198) to T.M.R. G.B.R. is a recipient of a postdoctoral fellowship award from the Ontario HIV Treatment network.

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  1. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, 01605, Massachusetts, USA

    G Brett Robb, Kirk M Brown, Jaspreet Khurana & Tariq M Rana

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Correspondence to Tariq M Rana.

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Robb, G., Brown, K., Khurana, J. et al. Specific and potent RNAi in the nucleus of human cells. Nat Struct Mol Biol 12, 133–137 (2005). https://doi.org/10.1038/nsmb886

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