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

  • Michael T. Massengill
  • Brianna M. Young
  • Alfred S. Lewin
  • Cristhian J. IldefonsoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1937)


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.

Key words

Gene therapy Recombinant adeno-associated virus Autosomal-dominant Retinal degeneration Allele-independent Short-hairpin RNA shRNA-resistant cDNA 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Michael T. Massengill
    • 1
  • Brianna M. Young
    • 2
  • Alfred S. Lewin
    • 1
    • 2
  • Cristhian J. Ildefonso
    • 1
    • 2
    Email author
  1. 1.Department of Molecular Genetics and MicrobiologyUniversity of Florida College of MedicineGainesvilleUSA
  2. 2.Department of OphthalmologyUniversity of Florida College of MedicineGainesvilleUSA

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