Intein-mediated protein trans-splicing expands adeno-associated virus transfer capacity in the retina
Creators
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Tornabene, Patrizia1
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Trapani, Ivana2
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Minopoli, Renato1
- Centrulo, Miriam1
- Lupo, Mariangela1
- de Simone, Sonia1
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Tiberi, Paola1
- Dell'Aquila, Fabio1
- Marrocco, Elena1
- Iodice, Carolina1
- Iuliano, Antonella1
- Gesualdo, Carlo3
- Rossi, Settimio3
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Giaquinto, Laura1
- Albert, Silvia4
- Hoyng, Carel B.4
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Polishchuk, Elena1
- Cremers, Frans P.M.5
- Surace, Enrico M.6
- Simonelli, Francesca3
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De Matteis, Maria A.7
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Polishchuk, Roman1
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Auricchio, Alberto8
- 1. Telethon Institute of Genetics and Medicine (TIGEM), 80078 Pozzuoli, Italy.
- 2. Telethon Institute of Genetics and Medicine (TIGEM), 80078 Pozzuoli, Italy - Medical Genetics, Department of Translational Medicine, Federico II University, 80131 Naples, Italy.
- 3. Eye Clinic, Multidisciplinary Department of Medical, Surgical and Dental Sciences, University of Campania L. Vanvitelli, 80131 Naples, Italy.
- 4. Department of Human Genetics and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525 Nijmegen, Netherlands.
- 5. Department of Ophthalmology and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525 Nijmegen, Netherlands.
- 6. Telethon Institute of Genetics and Medicine (TIGEM), 80078 Pozzuoli, Italy. - Medical Genetics, Department of Translational Medicine, Federico II University, 80131 Naples, Italy.
- 7. Telethon Institute of Genetics and Medicine (TIGEM), 80078 Pozzuoli, Italy. Department of Molecular Medicine and Medical Biotechnology, Federico II University, 80131 Naples, Italy.
- 8. Telethon Institute of Genetics and Medicine (TIGEM), 80078 Pozzuoli, Italy. - Department of Advanced Biomedicine, Federico II University, 80131 Naples, Italy.
Description
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.
Files
Tornabene et al 2019.pdf
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