June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Protein-based nanoparticles for non-viral retinal gene therapy.
Author Affiliations & Notes
  • Anna Salas Torras
    Ophthalmology, Vall d'Hebron Research Inst, Barcelona, Spain
  • Laura Fontrodona Montals
    Ophthalmology, Vall d'Hebron Research Inst, Barcelona, Spain
  • Neus Ferrer Miralles
    Universitat Autònoma de Barcelona, Bellaterra, Spain
  • Ibane Abasolo
    Vall d'Hebron Research Institute, Barcelona, Spain
  • Miguel A Zapata
    Ophthalmology, Vall d'Hebron Research Inst, Barcelona, Spain
  • Jose Garcia-Arumi
    Ophthalmology, Vall d'Hebron Research Inst, Barcelona, Spain
  • Footnotes
    Commercial Relationships Anna Salas Torras, None; Laura Fontrodona Montals, None; Neus Ferrer Miralles, None; Ibane Abasolo, None; Miguel Zapata, None; Jose Garcia-Arumi, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1537. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Anna Salas Torras, Laura Fontrodona Montals, Neus Ferrer Miralles, Ibane Abasolo, Miguel A Zapata, Jose Garcia-Arumi, ; Protein-based nanoparticles for non-viral retinal gene therapy. . Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1537.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: Protein-based nanoparticles for DNA compactation have been widely studied for different applications. In this work two different peptidic constructions (R9-GFP-His and HNRK) have been tested on in-vitro and in-vivo models for its application in non-viral retinal gene therapy.

Methods: R9-GFP-His and HNRK nanoparticles have been produced in bacteria and purified. Plasmidic DNA compactation efficacy has been studied by agarose gel electrophoresis assay. Transfection efficiency of both nanoparticles using a plasmid encoding tdTomato fluorescent protein has been first evaluated in-vitro in two cell lines, and therefore the best formulation has been used for in-vivo transfection assays by sub-retinal injections in Long-Evans rats. In vivo transfection efficiencies have been evaluated by direct fluorescence observation in cryosections and flat-mounts.

Results: R9-GFP-His and HNRK nanoparticles induce DNA condensation, producing a retardation in agarose gel electrophoresis. The in-vitro transfection with ptdTomato-NP complexes at different ratios showed lower values compared with the commercial reactive used as a positive control, being the highest NP-DNA ratios the most efficient. In rat retinas, sub-retinal administration transfected cells mainly in the RPE layer.

Conclusions: This study establishes the base for the application of these non-viral transfection systems for future treatments of retinal disorders.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×