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Gene transfer engineering for astrocyte-specific silencing in the CNS

Gene Therapy volume 22, pages 830–839 (2015)Cite this article

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Abstract

Cell-type-specific gene silencing is critical to understand cell functions in normal and pathological conditions, in particular in the brain where strong cellular heterogeneity exists. Molecular engineering of lentiviral vectors has been widely used to express genes of interest specifically in neurons or astrocytes. However, we show that these strategies are not suitable for astrocyte-specific gene silencing due to the processing of small hairpin RNA (shRNA) in a cell. Here we develop an indirect method based on a tetracycline-regulated system to fully restrict shRNA expression to astrocytes. The combination of Mokola-G envelope pseudotyping, glutamine synthetase promoter and two distinct microRNA target sequences provides a powerful tool for efficient and cell-type-specific gene silencing in the central nervous system. We anticipate our vector will be a potent and versatile system to improve the targeting of cell populations for fundamental as well as therapeutic applications.

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Acknowledgements

This work was partially supported by the CEA and the European Community’s Seventh Framework Program FP7/2007–2013 under the grant agreement number HEALTH-F5-2008-222925 (NEUGENE) and by the Swiss National Science Foundation 31003A-140945. We thank Carole Malgorn (CEA, France) and Lydie Boussicault (CEA, France) for their help in genotyping mice; Fanny Petit (CEA, France), Martine Guillermier (CEA, France), Diane Houitte (CEA, France), Virginie Zimmer (LCMN, Lausanne) and Alexia Spoerl (LCMN, Lausanne) for help with animal experiments; and Dr Emmanuel Brouillet (CEA, France) and the Cellular Imaging Facility (CIF, Lausanne) for their advice about microscopic acquisitions.

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Author notes

  1. N Merienne and A Delzor: These authors contributed equally to these work.

Authors and Affiliations

  1. Department of Clinical Neurosciences (DNC), Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital (CHUV), Lausanne, Switzerland

    N Merienne, A Viret, M Rey & N Déglon

  2. LCMN, Neuroscience Research Center (CRN), Lausanne University Hospital, Lausanne, Switzerland

    N Merienne, A Viret, M Rey & N Déglon

  3. Commissariat à l’Energie Atomique et aux Energies Alternatives (CEA), Institute of Biomedical Imaging (I2BM), Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France

    A Delzor, N Dufour & P Hantraye

  4. CNRS-CEA URA2210, Fontenay-aux-Roses, France

    A Delzor, N Dufour & P Hantraye

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  1. N Merienne
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Correspondence to N Déglon.

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Merienne, N., Delzor, A., Viret, A. et al. Gene transfer engineering for astrocyte-specific silencing in the CNS. Gene Ther 22, 830–839 (2015). https://doi.org/10.1038/gt.2015.54

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