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DNA Delivery in Adult Mouse Eyes: An Update with Corneal Outcomes

  • John M. Nickerson
  • Shannon E. Getz
  • Jana T. Sellers
  • Micah A. Chrenek
  • Penny Goodman
  • Christiana J. Bernal
  • Jeffrey H. Boatright
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1121)

Abstract

Ocular injection (intravitreal, subretinal, or into the anterior space) is an efficient approach to deliver many classes of drugs, cells, and other treatments to various cell types of the eye. In particular, subretinal injection is efficient since delivered agents accumulate as there is no dilution due to transport processes or diffusion and the volume of the interphotoreceptor space (IPS) is minimal (10–20 μl in the human eye, less than 1 μl in the mouse eye). We previously reported methods using subretinal injection and electroporation to deliver DNA to photoreceptor and retinal pigment epithelium cells in retinas of live mice (Johnson et al., 14:2211–2226; Nickerson et al. 884:53–69, 2012; Andrieu-Soler et al. 13:692–706, 2007). Here we detail further optimization of that approach and additionally report its use in delivering DNA expression plasmids to the corneal endothelium.

Key words

Subretinal Injection Electroporation Interphotoreceptor space Transfection Dilation Sclera Cornea Reporter gene expression Subretinal bleb Cornea Corneal endothelium 

Notes

Acknowledgments

This work was supported by the National Eye Institute (R01EY016470, R01EY021592, R01EY014026, P30EY006360, R24EY017045, T32EY007092), an unrestricted grant to the Department of Ophthalmology at Emory University from Research to Prevent Blindness, Inc., and the Katz Foundation.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • John M. Nickerson
    • 1
  • Shannon E. Getz
    • 1
  • Jana T. Sellers
    • 1
  • Micah A. Chrenek
    • 1
  • Penny Goodman
    • 1
  • Christiana J. Bernal
    • 1
  • Jeffrey H. Boatright
    • 1
  1. 1.Department of OphthalmologyEmory UniversityAtlantaUSA

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