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Updated: May 24, 2026
In vivo Electroporation of Developing Mouse Retina
Published on: June 24, 2011
Direct gene transfer with compacted DNA nanoparticles in retinal pigment epithelial cells: expression, repeat
Zongchao Han1, Adarsha Koirala, Rasha Makkia
1Department of Cell Biology, University of Oklahoma Health Sciences Center, BMSB 781, 940 Stanton L Young Blvd, Oklahoma City, OK 73104, USA.
Aim:
To evaluate the safety of compacted DNA nanoparticles (NPs) in retinal pigment epithelial (RPE) cells.
Materials & Methods:
Enhanced GFP expression cassettes controlled by the RPE-specific vitelloform macular dystrophy promoter were constructed with and without a bacterial backbone and compacted into NPs formulated with polyethylene glycol-substituted lysine 30-mers. Single or double subretinal injections were administered in adult BALB/c mice. Expression levels of enhanced GFP, proinflammatory cytokines and neutrophil/macrophage mediators, and retinal function by electroretinogram were evaluated at different time-points postinjection.
Results:
Immunohistochemistry and real-time PCR demonstrated that NPs specifically transfect RPE cells at a higher efficiency than naked DNA and similar results were observed after the second injection. At 6 h postinjections, a transient inflammatory response was observed in all cohorts, including saline, indicating an adverse effect to the injection procedure. Subsequently, no inflammation was detected in all experimental groups.
Conclusion:
This study demonstrates the safety and efficacy of NP-mediated RPE gene transfer therapy following multiple subretinal administrations.
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