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In vivo adenovirus-mediated gene transfer to newborn rat retinal pigment epithelial cells
Affiliation:1. Center for Human Genome Variation, Duke University School of Medicine, Durham, North Carolina, USA;2. Department of Medicine, University of Melbourne, Austin Health and Royal Melbourne Hospital, Melbourne, Australia;3. Edmond and Lily Safra Children’s Hospital, Sheba Medical Center, Ramat Gan, Israel;4. Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel;5. Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel;6. Department of Pediatrics, Duke University School of Medicine, Durham, North Carolina, USA;7. Pinchas Borenstein Talpiot Medical Leadership Program, Pediatric Neurology Unit, Chaim Sheba Medical Center, Tel HaShomer, Israel;8. Metabolic Disease Unit, Edmond and Lily Children’s Hospital, Sheba Medical Center, Ramat Gan, Israel;9. Department of Neurobiology, Duke University, Durham, North Carolina, USA;10. Danek Gertner Institute of Human Genetics, Sheba Medical Center, Ramat Gan, Israel;11. UCB NewMedicines, Slough, UK;12. Division of Brain Sciences, Department of Medicine, Imperial College London, London, UK;13. Present address: Department of Human Genetics, McGill University, Montreal, Quebec, Canada;14. Department of Human Genetics, Graduate School of Medicine, University of Tokyo, Tokyo, Japan
Abstract:A successful surgical access to the subretinal space is critical for achieving adenovirus-mediated gene transfer to the retinal pigment epithelial (RPE) cells or photoreceptor cells. We report a novel surgical approach allowing an efficient delivery of recombinant replication-deficient adenoviral vectors into the subretinal space of newborn rats. Our data suggest that this method may be useful for infecting reproducibly large areas of the RPE cell layer of normal newborn rats and should be applicable to RCS pups. We also show the feasibility of infecting ex vivo RPE cells in culture using the same recombinant adenoviral vector.
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