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Suppression of rds Expression by siRNA and Gene Replacement Strategies for Gene Therapy Using rAAV Vector

  • Hilda Petrs-Silva
  • Douglas Yasumura
  • Michael T. Matthes
  • Matthew M. LaVail
  • Alfred S. Lewin
  • William W. Hauswirth
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 723)

Abstract

Small interfering RNA (siRNA) is a promising tool for the treatment of dominant diseases. Autosomal dominant eye diseases like retinitis pigmentosa are a leading cause of blindness. Mutations in rds/peripherin lead to the degeneration of photoreceptors and are associated with several autosomal retinal diseases. Our goal is to develop a gene therapy for rds mutations. We describe an siRNA-based mutation-independent approach, targeting rds in which levels of endogenous mutant and wild-type mRNA were reduced, and an siRNA-resistant version of rds gene was supplied simultaneously. siRNAs and resistant rds were delivered to the photoreceptors by recombinant adeno-associated virus (rAAV) vector through subretinal injections. The retinal phenotype was examined, both structurally and functionally at different time points after rAAV delivery. We demonstrate suppression of rds transcript by up to 50% with concomitant expression of replacement transcript in the retina of mice in vivo. These results validate the concept of suppression of rds and replacement strategies of gene therapy with rAAV vectors containing siRNA.

Keywords

rds/peripherin Retinitis pigmentosa Photoreceptor degeneration Gene therapy Adeno-associated viral vector siRNA Gene replacement 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Hilda Petrs-Silva
    • 1
  • Douglas Yasumura
    • 2
  • Michael T. Matthes
    • 2
  • Matthew M. LaVail
    • 2
  • Alfred S. Lewin
    • 1
    • 3
  • William W. Hauswirth
    • 1
  1. 1.Departments of OphthalmologyInstitute of Biophysics, CCS, UFRJRio de JaneiroBrazil
  2. 2.Beckman Vision CenterUniversity of California San FranciscoSan FranciscoUSA
  3. 3.Departments of Molecular Genetics and MicrobiologyUniversity of FloridaGainesvilleUSA

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