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Abstract #19151 Published in IGR 3-1
Induction of gene into the rabbit eye by iontophoresis: preliminary report
Asahara T; Shinomiya K; Naito T; Shiota HJapanese Journal of Ophthalmology 2001; 45: 31-39
PURPOSE: To assess the transfer of 6-carboxyfluorescein (6-FAM)-labelled phosphorothioate oligonucleotides (S-ODNs) into the ocular tissues, their stability, and possibility of injury to the ocular tissues. METHODS: S-ODNs (2 ml/eye) were transduced noninvasively into albino rabbit eyes. The iontophoresis group consisted of six rabbits (12 eyes); the control group consisted of two rabbits (four eyes) given eye drops containing S-ODNs. Aqueous humor and vitreous humor were collected after iontophoresis, subjected to electrophoresis with a fluorescence DNA sequencer, and analyzed by the Gene Scan program. Frozen sections, 10-μm thick, were prepared for observation under a fluorescence microscope. A plasmid, 4.7 kbp in size, that expresses green fluorescent protein (GFP) was induced into both eyes of nine rabbits by the same procedure. RESULTS: In the iontophoresis group, S-ODNs were detected in the anterior chamber five minutes after the start of electrophoresis and in the vitreous after ten minutes. These S-ODNs maintained the same length as at the initial synthesis. S-ODNs could also be detected in the posterior retina 20 minutes after electrophoresis. No evidence of degeneration or inflammation due to the above procedure was found in the ocular tissues. Fluorescence showing GFP gene expression was found in the cornea, anterior chamber angle, and ciliary subepithelial tissues. CONCLUSIONS: These findings show that iontophoresis is an effective method for inducing genes into the rabbit eye.
Dr T. Asahara, Department of Ophthalmology, University of Tokushima School of Medicine, Tokushima, Japan
Classification:
11.9 Gene therapy (Part of: 11 Medical treatment)