April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Pigs Transgenic For Human Dominant Mutations Of The GUCY2D Gene
Author Affiliations & Notes
  • Yvan Arsenijevic
    Unit of Gene Therapy & Stem Cell Biology, Jules-Gonin Eye Hospital, Univ Lausanne, Lausanne, Switzerland
  • Tim King
    Division of Developmental Biology, The Roslin Institute, Univ Edinburgh, Roslin, United Kingdom
  • Sylvain V. Crippa
    Unit of Gene Therapy & Stem Cell Biology, Jules-Gonin Eye Hospital, Univ Lausanne, Lausanne, Switzerland
  • Simon Lillico
    Division of Developmental Biology, The Roslin Institute, Univ Edinburgh, Roslin, United Kingdom
  • Chamsy Sarkis
    Team of Biotherapy and Biotechnology, CRICM, Paris, France
    NewVectys, Paris, France
  • Jacques Mallet
    Team of Biotherapy and Biotechnology, CRICM, Paris, France
  • Corinne Kostic
    Unit of Gene Therapy & Stem Cell Biology, Jules-Gonin Eye Hospital, Univ Lausanne, Lausanne, Switzerland
  • Bruce Whitelaw
    Division of Developmental Biology, The Roslin Institute, Univ Edinburgh, Roslin, United Kingdom
  • Footnotes
    Commercial Relationships  Yvan Arsenijevic, FAA & Univ Lausanne, Switzerland (P); Tim King, None; Sylvain V. Crippa, None; Simon Lillico, None; Chamsy Sarkis, CNRS & NewVectys, France (P), NewVectys, Paris, France (E); Jacques Mallet, None; Corinne Kostic, FAA, Univ Lausanne, Switzerland (P); Bruce Whitelaw, Univ Edinburgh, UK (P)
  • Footnotes
    Support  FP6 INTEGRA 029025-2, Retina France
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4340. doi:
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      Yvan Arsenijevic, Tim King, Sylvain V. Crippa, Simon Lillico, Chamsy Sarkis, Jacques Mallet, Corinne Kostic, Bruce Whitelaw; Pigs Transgenic For Human Dominant Mutations Of The GUCY2D Gene. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4340.

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Abstract

Purpose: : Studies on large animal models are an important step to test new therapeutical strategies before human application. Considering the importance of cone function for human vision and the paucity of large animal models for cone dystrophies having an enriched cone region, we propose to develop a pig model for cone degeneration. With a lentiviral-directed transgenesis, we obtained pigs transgenic for a cone-dominant mutant gene described in a human cone dystrophy.

Methods: : Lentiviral vectors encoding the human double mutant GUCY2DE837D/R838S cDNA under the control of a region of the pig arrestin-3 promoter (Arr3) was produced and used for lentiviral-derived transgenesis in pigs. PCR-genotyping and southern blotting determined the genotype of pigs born after injection of the vector at the zygote stage. Retina function analysis was performed by ERG and behavioral tests at 11, 24 and 54 weeks of age. OCT and histological analyses were performed to describe the retina morphology.

Results: : The ratio of transgenic pigs born after lentiviral-directed transgenesis was close to 50%. Transgenic pigs with 3 to 5 transgene copies per cell clearly present a reduced photopic response from 3 months of age on. Except for one pig, which has 6 integrated transgene copies, no dramatic decrease in general mobility was observed even at 6 months of age. OCT examinations reveal no major changes in the ONL structure of the 6-months old pigs. The retina morphology was well conserved in the 2 pigs sacrificed (3 and 6 months old) except a noticeable displacement of some cone nuclei in the outer segment layer.

Conclusions: : Lentiviral-directed transgenesis is a rapid and straightforward method to engineer transgenic pigs. Some Arr3-GUCY2DE837D/R838S pigs show signs of retinal dysfunction but further work is needed to describe the progression of the disease in this model.

Keywords: retinitis • photoreceptors • genetics 
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