April 2011
Volume 52, Issue 14
ARVO Annual Meeting Abstract  |   April 2011
Mice Transgenic For Human Dominant Mutations Of The GUCY2D Gene
Author Affiliations & Notes
  • Corinne Kostic
    Gene Therapy & Stem Cell Biol, Jules-Gonin Eye Hosp, Univ Lausanne, Lausanne, Switzerland
  • Vérène Pignat
    Gene Therapy & Stem Cell Biol, Jules-Gonin Eye Hosp, Univ Lausanne, Lausanne, Switzerland
  • Bruce Whitelaw
    Division of Developmental Biology, The Roslin Institute, Univ of Edinburgh, Roslin, United Kingdom
  • Héloise Pilet
    NewVectys, Paris, France
  • Suzanne Ursulet
    NewVectys, Paris, France
  • Sébastien Dussaud
    Centre d'Expérimentation Fonctionnelle, Faculté de Médecine Pierre Marie Curie, Paris, France
  • Yvan Arsenijevic
    Gene Therapy & Stem Cell Biol, Jules-Gonin Eye Hosp, Univ Lausanne, Lausanne, Switzerland
  • Chamsy Sarkis
    NewVectys, Paris, France
    Team of Biotherapy and Biotechnology, CRICM, Paris, France
  • Footnotes
    Commercial Relationships  Corinne Kostic, FAA & Univ Lausanne, Switzerland (P); Vérène Pignat, None; Bruce Whitelaw, Univ Edinburgh, UK (P); Héloise Pilet, NewVectys, Paris, France (E); Suzanne Ursulet, NewVectys, Paris, France (E); Sébastien Dussaud, None; Yvan Arsenijevic, FAA & Univ Lausanne, Switzerland (P); Chamsy Sarkis, CNRS & NewVectys, France (P), NewVectys, Paris, France (E)
  • Footnotes
    Support  FP6 INTEGRA 029025-2
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4343. doi:
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      Corinne Kostic, Vérène Pignat, Bruce Whitelaw, Héloise Pilet, Suzanne Ursulet, Sébastien Dussaud, Yvan Arsenijevic, Chamsy Sarkis; Mice Transgenic For Human Dominant Mutations Of The GUCY2D Gene. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4343.

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

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Purpose: : Animal models are essential to study pathological mechanisms and to test new therapeutic strategies. Many mouse models mimic human rod loss but only a limited number simulate cone dystrophies. The importance of cone function for human vision highlights the need to engineer a model for cone degeneration. An approach of lentiviral-directed transgenesis was tested in mice to express a dominant mutant gene described in a human cone dystrophy.

Methods: : Lentiviral vectors (LV) encoding either hrGFPII or the human double mutant GUCY2DE837D/R838S cDNA under the control of a region of the pig arrestin-3 promoter (Arr3) were produced and used for lentiviral-derived transgenesis. PCR-genotyping determined the transgenic mouse ratio. The expression of GFP was then analyzed both in vivo and by immunohistochemistry in Arr3-GFPII mice. Functional analysis was performed by ERG at 5, 9, 16 and 24 weeks for Arr3-GUCY2DE837D/R838S mice. Mice were sacrificed at 10 months of age for both histological analysis and RNA extraction.

Results: : While all the newborns from the transgenesis using the LV-Arr3-GFPII were transgenic, one third of the newborns from the LV-Arr3-GUCY2DE837D/R838S transgenesis were positive. Expression of GFPII was demonstrated by in vivo imaging, while expression of the mutant GUCY2D transcript was detetected using RT-PCR. No severe alteration of the functional response was observed up to 24 weeks of age in the transgenic mice. No obvious modification of the retinal morphology was identified either.

Conclusions: : Lentiviral-directed transgenesis is a rapid and straightforward method to engineer transgenic mice. Protein expression can be specifically targeted to the retina and thus could help to study the effect of expression of dominant mutant proteins. In our case, Arr3-GUCY2DE837D/R838S mice have a less severe phenotype than that described for human patients. Further analyses are required to understand this difference but several modifications of the expression cassette might also help to increase the expression of the mutant protein and reinforce the phenotype. Interestingly, the same construct is less effective in mouse versus pig retina (see Arsenijevic et al. ARVO 2011 abstract).

Keywords: retinitis • photoreceptors • genetics 

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