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In Vivo Electroporation of Developing Mouse Retina

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Retinal Gene Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1715))

Abstract

In vivo electroporation enables the transformation of retinal tissue with engineered DNA plasmids, facilitating the selective expression of desired gene products. This method achieves plasmid transfer via the application of an external electrical field, which both generates a transient increase in the permeability of cell plasma membranes, and promotes the incorporation of DNA plasmids by electrophoretic transfer through the permeabilized membranes. Here we describe a method for the preparation, injection, and electroporation of DNA plasmids into neonatal mouse retinal tissue. This method can be utilized to perform gain of function or loss of function studies in the mouse. Experimental design is limited only by construct availability.

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Author information

Authors and Affiliations

  1. Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Jimmy de Melo & Seth Blackshaw

  2. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  3. Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  4. Center for Human Systems Biology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  5. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

Authors
  1. Jimmy de Melo
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  2. Seth Blackshaw
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Corresponding author

Correspondence to Seth Blackshaw .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology, Leiden University Medical Center (LUMC), Leiden, The Netherlands

    Camiel J.F. Boon

  2. Department of Ophthalmology, Leiden University Medical Center (LUMC), Leiden, The Netherlands

    Jan Wijnholds

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de Melo, J., Blackshaw, S. (2018). In Vivo Electroporation of Developing Mouse Retina. In: Boon, C., Wijnholds, J. (eds) Retinal Gene Therapy. Methods in Molecular Biology, vol 1715. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7522-8_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7522-8_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7521-1

  • Online ISBN: 978-1-4939-7522-8

  • eBook Packages: Springer Protocols

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