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Neuron-restrictive silencer elements mediate neuron specificity of adenoviral gene expression

Nature Biotechnology volume 17, pages 865–869 (1999)Cite this article

Abstract

Neuron–restrictive silencer elements (NRSEs) were used to target the gene expression of adenoviral vectors specifically to neuron cells in the central nervous system. By generating adenoviral constructs in which NRSE sequences were placed upstream from the ubiquitous phosphoglycerate kinase promoter, the specificity of expression of a luciferase reporter gene was tested in both cell lines and primary cultures. Whereas transgene expression was negligible in nonneuronal cells following infection with an adenovirus containing 12 NRSEs, neuronal cells strongly expressed luciferase when infected with the same adenovirus. The NRSEs restricted expression of the luciferase gene to neuronal cells in vivo when adenoviruses were injected both intramuscularly into mice and intracerebrally into rats. This NRSE strategy may avoid side effects resulting from the ectopic expression of therapeutic genes in the treatment of neurological diseases. In particular, it may allow the direct transfection of motor neurons without promoting transgene expression within inoculated muscles or the secretion of transgene products into the bloodstream.

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Figure 1: Transfection of adenoviral plasmids.
Figure 2: Transfection of NSE and NRSE plasmids.
Figure 3: Adenoviral infection in vitro of cell lines (A, B) and rat primary cultures (C–F).
Figure 4: Detection of luciferase-expressing neurons in the brainstem one week after adenovirus injection into the tongue.
Figure 5: Luciferase activity after adenoviral in vivo infection.
Figure 6: Double immunostaining of luciferase/NeuN neurons and luciferase/GFAP astrocytes after injection of Ad-N12PGK-luc and Ad-PGK-luc into the rat striatum (STR, A–D) and hippocampus (HPC, E–H).

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Acknowledgements

The authors thank Véronique Albanese, Fabienne Chatail, and Sylvie Dumas for helpful suggestions, and Sylvie Carpaille and Philippe Colin for handling the animals. S.M. was supported by Ministère de l'Education Nationale de l'Enseignement Supérieur et de la Recherche, H.K. by Institut de Formation Supérieure Biomédicale, and M.B. by the European Commission Biotechnology program. This work was supported in part by Association Française contre les Myopathies, Association Française Retinis Pigmentosa, Institut de Recherche sur la Moelle Epinière, Centre National de la Recherche Scientifique, Rhône-Poulenc Rorer, and the Conseil Régional d'Ile-de-France.

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  1. Laboratoire de Génétique Moléculaire de la Neurotransmission et des Processus Neurodégénératifs, CNRS UMR C9923, Hôpital de la Pitié Salpêtrière , Paris, France

    Stéphanie Millecamps, Hélène Kiefer, Vincent Navarro, Marie-Claude Geoffroy, Jean-Jacques Robert, Jacques Mallet & Martine Barkats

  2. Rhône-Poulenc Rorer, Vitry sur seine , France

    Jean-Jacques Robert & Françoise Finiels

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  1. Stéphanie Millecamps
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Correspondence to Jacques Mallet.

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Millecamps, S., Kiefer, H., Navarro, V. et al. Neuron-restrictive silencer elements mediate neuron specificity of adenoviral gene expression. Nat Biotechnol 17, 865–869 (1999). https://doi.org/10.1038/12849

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