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Exogenous control of mammalian gene expression via modulation of translational termination

Nature Medicine volume 12, pages 1093–1099 (2006)Cite this article

Abstract

Here, we describe a system for the exogenous control of gene expression in mammalian cells that relies on the control of translational termination. To achieve gene regulation, we modified protein-coding sequences by introduction of a translational termination codon just downstream from the initiator AUG codon. Translation of the resulting mRNA leads to potent reduction in expression of the desired gene product. Expression of the gene product can be controlled by treating cells that express the mRNA with either aminoglycoside antibiotics or several nonantibiotic compounds. We show that the extent of regulation of gene expression can be substantial (60-fold) and that regulation can be achieved in the case of a variety of different genes, in different cultured cell lines and in primary cells in vivo. This gene regulation strategy offers significant advantages over existing methods for controlling gene expression and should have both immediate experimental application and possible clinical application.

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Figure 1: Design features of translation-based system for gene regulation.
Figure 2: Aminoglycoside-induced suppression of translational termination can be used to regulate gene expression in cultured cells.
Figure 3: Other aminoglycosides and nonantibiotic compounds can be used to regulate gene expression via suppression of nonsense mutations.
Figure 4: In vivo regulation of gene expression in mouse lungs and mouse hematopoietic cells.

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Acknowledgements

This work was supported by the National Institutes of Health grant 5PO-HL54785 (to R.C.M.) and by a grant from L'Association Francaise contre les Myopathies (AFM). We thank J. Mulligan for preparation of the artwork used in the figures, P. Russell for technical assistance and A. Balazs for helpful discussions. We particularly acknowledge early discussions with J. Leiden and L. Sweeney regarding the possibility of applying aminoglycoside suppression of nonsense codons to the development of gene regulation systems.

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  1. Department of Genetics, and Division of Molecular Medicine, Harvard Medical School, Children's Hospital, Boston, 02115, Massachusetts, USA

    George J Murphy, Gustavo Mostoslavsky, Darrell N Kotton & Richard C Mulligan

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  1. George J Murphy
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Correspondence to Richard C Mulligan.

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Murphy, G., Mostoslavsky, G., Kotton, D. et al. Exogenous control of mammalian gene expression via modulation of translational termination. Nat Med 12, 1093–1099 (2006). https://doi.org/10.1038/nm1376

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