May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Targeting Gene Expression to the Trabecular Meshwork (TM) in the Rat Eye With the Promoter of a Member of the Alpha 2u-Globulin Family
Author Affiliations & Notes
  • P. Gonzalez
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • C. Luna
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • P. Challa
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • P. B. Liton
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • G. Li
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • J. Qiu
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • D. L. Epstein
    Ophthalmology, Duke University Eye Center, Durham, North Carolina
  • Footnotes
    Commercial Relationships  P. Gonzalez, None; C. Luna, None; P. Challa, None; P.B. Liton, None; G. Li, None; J. Qiu, None; D.L. Epstein, None.
  • Footnotes
    Support  NIH Grants EY01894,EY016228,EY05722, American Health Assistance Foundation, and Research to Prevent Blindness
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 1623. doi:
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      P. Gonzalez, C. Luna, P. Challa, P. B. Liton, G. Li, J. Qiu, D. L. Epstein; Targeting Gene Expression to the Trabecular Meshwork (TM) in the Rat Eye With the Promoter of a Member of the Alpha 2u-Globulin Family. Invest. Ophthalmol. Vis. Sci. 2008;49(13):1623.

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

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Abstract

Purpose: : To test the potential of the promoter of the alpha 2u-globulin gene to target gene expression to the trabecular meshwork (TM) in the rat eye in vivo.

Methods: : An 800 bp fragment of the promoter from a member of the alpha 2u-globulin family identified by gene array analysis was generated by genome walking PCR from rat genomic DNA. An adenoviral vector expressing Cre recombinase under this promoter fragment (Ad. Alpha 2u-cre) and a second adenoviral vector including the CMV promoter, a loxed stop sequence, and the reporter gene EGFP (Ad.CMV-stop-EGFP), were co-injected in the anterior chamber of living rat eyes at 5x106 to 107 pfus. Control rat eyes were injected with the same pfus of an adenovirus expressing Cre under the CMV promoter (Ad.CMV-cre) and the same reporter adenovirus as the experimental eyes. The pattern of expression of EGFP in the eye was analyzed in cryosections as well as in semi-thin sections.

Results: : Rat eyes injected with the Ad.Alpha 2u-cre and Ad.CMV-stop-EGFP viral constructs demonstrated preferential expression of the reporter gene EGFP in the cells of the TM compared to the controls injected with the Ad.CMV-cre and the Ad.CMV-stop-EGFP viruses.

Conclusions: : The 800 bp promoter fragment from the alpha 2u-globulin gene should be useful to target transgene expression to the TM in vivo using viral vectors injected into the anterior chamber. Current efforts are been made to test the potential of this promoter for targeting gene expression to the TM in transgenic mice. The availability of such a promoter would allow the creation of new animal models relevant to glaucoma research.

Keywords: trabecular meshwork • gene/expression • adenovirus 
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