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Co-Delivery of a Short-Hairpin RNA and a shRNA-Resistant Replacement Gene with Adeno-Associated Virus: An Allele-Independent Strategy for Autosomal-Dominant Retinal Disorders

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Viral Vectors for Gene Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1937))

Abstract

Recombinant adeno-associated virus (rAAV) has become an important gene delivery vector for the treatment of inherited retinal degenerative diseases. Many of the mutations leading to retinal degeneration are inherited in an autosomal-dominant pattern and can produce toxic gain-of-function and/or dominant-negative effects. Here we describe an allele-independent gene therapy strategy with rAAV to treat autosomal-dominant retinal degenerative diseases. In this methodology, we co-deliver a short-hairpin RNA (shRNA) to inhibit expression of both the toxic and (WT) copies of the gene as well as an shRNA-resistant cDNA for functional gene replacement with a rAAV.

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Acknowledgments

This work was funded by an F30 from the NEI (MM), an R01 from the NEI (ASL), a grant from the Bright Focus Foundation (CJI) and an unrestricted grant from the Research to Prevent Blindness (CJI).

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Authors and Affiliations

  1. Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL, USA

    Michael T. Massengill, Alfred S. Lewin & Cristhian J. Ildefonso

  2. Department of Ophthalmology, University of Florida College of Medicine, Gainesville, FL, USA

    Brianna M. Young, Alfred S. Lewin & Cristhian J. Ildefonso

Authors
  1. Michael T. Massengill
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  2. Brianna M. Young
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  3. Alfred S. Lewin
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  4. Cristhian J. Ildefonso
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Corresponding author

Correspondence to Cristhian J. Ildefonso .

Editor information

Editors and Affiliations

  1. Translational Science and Molecular Medicine, Michigan State University, Grand Rapids, MI, USA

    Fredric P. Manfredsson

  2. Translational Science and Molecular Medicine, Michigan State University, Grand Rapids, MI, USA

    Matthew J. Benskey

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Massengill, M.T., Young, B.M., Lewin, A.S., Ildefonso, C.J. (2019). Co-Delivery of a Short-Hairpin RNA and a shRNA-Resistant Replacement Gene with Adeno-Associated Virus: An Allele-Independent Strategy for Autosomal-Dominant Retinal Disorders. In: Manfredsson, F., Benskey, M. (eds) Viral Vectors for Gene Therapy. Methods in Molecular Biology, vol 1937. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9065-8_15

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  • DOI: https://doi.org/10.1007/978-1-4939-9065-8_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9064-1

  • Online ISBN: 978-1-4939-9065-8

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