Our research objective is to use DNA nanotechnology and helper-independent Sleeping Beauty Trasposon-Transposase motif to develop a novel and effective therapeutic strategy for eye diseases; particularly those associated with loss-of-function mutations in the retina and retinal pigment epithelium (RPE). Successful application of ocular therapy is contingent upon the efficient delivery and targeting of therapeutic agents in a cell-specific manner. We propose to test the hypothesis that compacted DNA nanoparticles is an effective, efficient, and well-tolerated method for therapeutic gene delivery and targeting to the retina and RPE cells to respectively battle diseases of the retina and RPE. Since nanoparticles are small and condensed, they can pass safely through the cell membrane as well as the nuclear membrane and into the nucleus. Our experimental plan requires three components: engineered nanparticles, genetically engineered therapy, and animal models of ocular diseases, all of which are currently available in our laboratory. Therapeutic rescue of retinal disease will be assessed functionally using electroretinography (ERG) and multifocal ERG, structural analysis will be assessed using immunofluorescence, light and electron microscopy, and biochemical analysis will use real time RT-PCR, RNase protection, Western blot, and ELISA analysis. When taken together, these studies will provide valuable insight into non-viral gene therapy as a prospective method for the treatment of inherited blinding diseases. ? ? ?
Elliott, Michael H; Nash, Zack A; Takemori, Nobuaki et al. (2008) Differential distribution of proteins and lipids in detergent-resistant and detergent-soluble domains in rod outer segment plasma membranes and disks. J Neurochem 104:336-52 |
Chakraborty, Dibyendu; Ding, Xi-Qin; Fliesler, Steven J et al. (2008) Outer segment oligomerization of Rds: evidence from mouse models and subcellular fractionation. Biochemistry 47:1144-56 |
Cai, Xue; Conley, Shannon; Naash, Muna (2008) Nanoparticle applications in ocular gene therapy. Vision Res 48:319-24 |
Nour, May; Fliesler, Steven J; Naash, Muna I (2008) Genetic supplementation of RDS alleviates a loss-of-function phenotype in C214S model of retinitis pigmentosa. Adv Exp Med Biol 613:129-38 |
Farjo, Rafal; Skaggs, Jeff; Quiambao, Alexander B et al. (2006) Efficient non-viral ocular gene transfer with compacted DNA nanoparticles. PLoS One 1:e38 |