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Visualization and selective chemical targeting of RNA G-quadruplex structures in the cytoplasm of human cells

Nature Chemistry volume 6, pages 75–80 (2014)Cite this article

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Abstract

Following extensive evidence for the formation of four-stranded DNA G-quadruplex structures in vitro, DNA G-quadruplexes have been observed within human cells. Although chemically distinct, RNA can also fold in vitro into G-quadruplex structures that are highly stable because of the 2′-hydroxyl group. However, RNA G-quadruplexes have not yet been reported in cells. Here, we demonstrate the visualization of RNA G-quadruplex structures within the cytoplasm of human cells using a G-quadruplex structure-specific antibody. We also demonstrate that small molecules that bind to G-quadruplexes in vitro can trap endogenous RNA G-quadruplexes when applied to cells. Furthermore, a small molecule that exhibits a preference for RNA G-quadruplexes rather than DNA G-quadruplexes in biophysical experiments also shows the same selectivity within a cellular context. Our findings provide substantive evidence for RNA G-quadruplex formation in the human transcriptome, and corroborate the selectivity and application of stabilizing ligands that target G-quadruplexes within a cellular context.

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Figure 1: The G-quadruplex structure-specific antibody, BG4, binds with high affinity to RNA G-quadruplex structures.
Figure 2: Visualization of RNA G-quadruplexes in the cytoplasm of human cells.
Figure 3: RNA G-quadruplex structures in human cells are stabilized by a small-molecule G-quadruplex ligand.
Figure 4: Selective stabilization of endogenous RNA, but not DNA, G-quadruplexes after cell treatment with a RNA G-quadruplex-specific ligand.

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Acknowledgements

We thank D. Sanders for stimulating discussions and Cancer Research UK for funding.

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Authors and Affiliations

  1. Cancer Research UK Cambridge Institute, Li Ka Shing Centre, Robinson Way, Cambridge, CB2 0RE, UK

    Giulia Biffi, David Tannahill & Shankar Balasubramanian

  2. Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK

    Marco Di Antonio & Shankar Balasubramanian

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  1. Giulia Biffi
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  2. Marco Di Antonio
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Contributions

G.B. and M.D.A. carried out the experiments. G.B., M.D.A., D.T. and S.B. designed the experiments. G.B., D.T. and S.B. co-wrote the manuscript.

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Correspondence to Shankar Balasubramanian.

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The authors declare no competing financial interests.

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Biffi, G., Di Antonio, M., Tannahill, D. et al. Visualization and selective chemical targeting of RNA G-quadruplex structures in the cytoplasm of human cells. Nature Chem 6, 75–80 (2014). https://doi.org/10.1038/nchem.1805

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