efficient non-viral ocular gene transfer with compacted dna nanoparticles高效病毒性眼部基因转移与压实dna纳米颗粒.pdfVIP

Efficient Non-Viral Ocular Gene Transfer with Compacted DNA Nanoparticles 1 1 1 2 1 Rafal Farjo , Jeff Skaggs , Alexander B. Quiambao , Mark J. Cooper , Muna I. Naash * 1 Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States of America, 2 Copernicus Therapeutics, Inc., Cleveland, Ohio, United States of America Background. The eye is an excellent candidate for gene therapy as it is immune privileged and much of the disease-causing genetics are well understood. Towards this goal, we evaluated the efficiency of compacted DNA nanoparticles as a system for non-viral gene transfer to ocular tissues. The compacted DNA nanoparticles examined here have been shown to be safe and effective in a human clinical trial, have no theoretical limitation on plasmid size, do not provoke immune responses, and can be highly concentrated. Methods and Findings. Here we show that these nanoparticles can be targeted to different tissues within the eye by varying the site of injection. Almost all cell types of the eye were capable of transfection by the nanoparticle and produced robust levels of gene expression that were dose-dependent. Most impressively, subretinal delivery of these nanoparticles transfected nearly all of the photoreceptor population and produced expression levels almost equal to that of rod opsin, the highest expressed gene in the retina. Conclusions. As no deleterious effects on retinal function were observed, this treatment strategy appears to be clinically viable and provides a highly efficient non-viral technology to safely deliver and express nucleic acids in the retina and other ocular tissues. Citation: Farjo R, Skaggs J, Quiambao AB, Cooper MJ, Naash MI (2006) Efficient Non-Viral Ocular Gene Transfer with Compacted