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Recombinant Alphavirus-Mediated Expression of Ion Channels and Receptors in the Brain

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Receptor and Ion Channel Detection in the Brain

Part of the book series: Neuromethods ((NM,volume 169))

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Abstract

Ectopic gene transfer has been a crucial method for neurobiological studies aiming to resolve the function of specific membrane and non-membrane proteins. For this purpose, expression vectors based on Semliki Forest virus, Sindbis virus, and Venezuelan equine encephalitis virus, all members of the Alphavirus genus, have been engineered in the form of naked RNA replicons, recombinant viral particles, and layered DNA vectors. By applying these alphavirus vectors to mammalian cell lines, large quantities of integral membrane proteins including ligand- and voltage-gated ion channels as well as G protein-coupled receptors have been expressed, and their functional activity has been confirmed by both electrophysiological recordings and coupling to G proteins. Furthermore, alphavirus vectors have provided efficient delivery tools for high-level expression of recombinant proteins in cultures of primary neurons and organotypic hippocampal slices, as well as upon intracranial injection, in vivo in rodent brain. This chapter describes the generation of infectious but replication-deficient Semliki Forest virus and Sindbis virus particles that are useful for the efficient transduction of neurons in vitro, in situ, and in vivo.

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Acknowledgments

The authors thank Dr. Sondra Schlesinger for her comments on the manuscript.

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Authors and Affiliations

  1. Department of Biology, Kantonsschule Hohe Promenade, Zurich, Switzerland

    Markus U. Ehrengruber

  2. PanTherapeutics, Lutry, Switzerland

    Kenneth Lundstrom

Authors
  1. Markus U. Ehrengruber
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  2. Kenneth Lundstrom
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Corresponding author

Correspondence to Markus U. Ehrengruber .

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Editors and Affiliations

  1. Instituto de Investigación en Discapacidades Neurológicas (IDINE), Departamento de Ciencias Médicas, Facultad de Medicina, Universidad Castilla-La Mancha, Albacete, Spain

    Rafael Lujan

  2. Pharmacology Unit, Department of Pathology and Experimental Therapeutics, School of Medicine and Health Sciences, IDIBELL, Universitat de Barcelona, L’Hospitalet de Llobregat, Barcelona, Spain

    Francisco Ciruela

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© 2021 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ehrengruber, M.U., Lundstrom, K. (2021). Recombinant Alphavirus-Mediated Expression of Ion Channels and Receptors in the Brain. In: Lujan, R., Ciruela, F. (eds) Receptor and Ion Channel Detection in the Brain. Neuromethods, vol 169. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1522-5_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1522-5_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1521-8

  • Online ISBN: 978-1-0716-1522-5

  • eBook Packages: Springer Protocols

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