1. Intein-mediated protein trans-splicing expands adeno-associated virus transfer capacity in the retina.
- Author
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Tornabene P, Trapani I, Minopoli R, Centrulo M, Lupo M, de Simone S, Tiberi P, Dell'Aquila F, Marrocco E, Iodice C, Iuliano A, Gesualdo C, Rossi S, Giaquinto L, Albert S, Hoyng CB, Polishchuk E, Cremers FPM, Surace EM, Simonelli F, De Matteis MA, Polishchuk R, and Auricchio A
- Subjects
- Animals, Genetic Vectors administration & dosage, Genetic Vectors metabolism, Green Fluorescent Proteins metabolism, Humans, Induced Pluripotent Stem Cells metabolism, Mice, Organoids ultrastructure, Organoids virology, Phenotype, Photoreceptor Cells, Vertebrate metabolism, Photoreceptor Cells, Vertebrate virology, Swine, Dependovirus genetics, Gene Transfer Techniques, Green Fluorescent Proteins genetics, Inteins, Retina virology, Trans-Splicing genetics
- Abstract
Retinal gene therapy with adeno-associated viral (AAV) vectors holds promises for treating inherited and noninherited diseases of the eye. Although clinical data suggest that retinal gene therapy is safe and effective, delivery of large genes is hindered by the limited AAV cargo capacity. Protein trans-splicing mediated by split inteins is used by single-cell organisms to reconstitute proteins. Here, we show that delivery of multiple AAV vectors each encoding one of the fragments of target proteins flanked by short split inteins results in protein trans-splicing and full-length protein reconstitution in the retina of mice and pigs and in human retinal organoids. The reconstitution of large therapeutic proteins using this approach improved the phenotype of two mouse models of inherited retinal diseases. Our data support the use of split intein-mediated protein trans-splicing in combination with AAV subretinal delivery for gene therapy of inherited blindness due to mutations in large genes., (Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2019
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