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Intravitreal Injection of AAV for the Transduction of Mouse Retinal Ganglion Cells

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Retinal Ganglion Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2708))

Abstract

The injection of therapies into the eye is common practice, both clinically and pre-clinically. The most straightforward delivery route is via an intravitreal injection, which introduces the treatment into the largest cavity at the posterior of the eye. This technique is frequently used to deliver gene therapies, including those containing recombinant adeno-associated viral vectors (AAVs), to the back of the eye to enable inner retinal targeting. This chapter provides detailed methodology on how to successfully perform an intravitreal injection in mice. The chapter covers vector preparation considerations, advice on how to minimize vector loss in the injection device, and ways to reduce vector reflux from the eye when administering a therapy. Finally, a protocol is provided on common retinal histology processing techniques to assess vector-mediated expression in retinal ganglion cells. It is hoped that this chapter will enable researchers to carry out effective and consistent intravitreal injections that transduce the inner retinal surface while avoiding common pitfalls.

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Acknowledgments

Bart Nieuwenhuis is supported by the Medical Research Council (MR/V002694/1) in the form of a postdoctoral research associate position. Andrew Osborne is supported by Ikarovec Limited. We thank Andrea Loreto, Fiona Love, and Yamunadevi Lakshmanan for proofreading and Laura Vaux, Emily Warner, and Kara Boyd for anesthetic support and imaging.

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Authors and Affiliations

  1. John van Geest Centre for Brain Repair, Department of Clinical Neurosciences, University of Cambridge, Cambridge, UK

    Bart Nieuwenhuis & Andrew Osborne

  2. Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK

    Bart Nieuwenhuis

  3. Ikarovec Ltd, The Norwich Research Park Innovation Centre, Norwich, UK

    Andrew Osborne

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  1. Bart Nieuwenhuis
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  2. Andrew Osborne
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Correspondence to Andrew Osborne .

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Editors and Affiliations

  1. School of Optometry and Vision Sciences, Cardiff University, Cardiff, UK

    Ben Mead

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Nieuwenhuis, B., Osborne, A. (2023). Intravitreal Injection of AAV for the Transduction of Mouse Retinal Ganglion Cells. In: Mead, B. (eds) Retinal Ganglion Cells. Methods in Molecular Biology, vol 2708. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3409-7_17

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  • DOI: https://doi.org/10.1007/978-1-0716-3409-7_17

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