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Retinal Degeneration pp 59–74Cite as

CRISPR/Cas9 Gene Editing In Vitro and in Retinal Cells In Vivo

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1834))

Abstract

CRISPR/Cas9 is an efficient tool to knock down specific genes in various organisms. In this chapter, we describe how to assess knockdown of human rhodopsin (RHO) gene carrying the P23H mutation in vitro, in engineered HeLa cells, and in vivo, in P23H RHO transgenic mice. To this aim, we report two molecular assays: site-specific PCR on P23H RHO cells treated with CRISPR/Cas9 and Western blotting analysis on retinal cells prepared from P23H RHO transgenic mice electroporated with CRISPR/Cas9 and GFP plasmids.

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Acknowledgment

This work was supported by Fondazione Roma, Call for Retinitis Pigmentosa.

Author information

Authors and Affiliations

  1. Department of Life Sciences, Centre for Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy

    Daniela Benati & Alessandra Recchia

  2. Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy

    Valeria Marigo

Authors
  1. Daniela Benati
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  2. Valeria Marigo
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  3. Alessandra Recchia
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Corresponding author

Correspondence to Alessandra Recchia .

Editor information

Editors and Affiliations

  1. Institute of Human Genetics, University of Regensburg, Regensburg, Germany

    Bernhard H. F. Weber

  2. Department of Ophthalmology, University of Cologne, Cologne, Germany

    Thomas Langmann

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Benati, D., Marigo, V., Recchia, A. (2019). CRISPR/Cas9 Gene Editing In Vitro and in Retinal Cells In Vivo. In: Weber, B.H.F., Langmann, T. (eds) Retinal Degeneration. Methods in Molecular Biology, vol 1834. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8669-9_4

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  • DOI: https://doi.org/10.1007/978-1-4939-8669-9_4

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8668-2

  • Online ISBN: 978-1-4939-8669-9

  • eBook Packages: Springer Protocols

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