Intein-mediated protein trans-splicing expands adeno-associated virus transfer capacity in the retina.
Animals
Dependovirus
/ genetics
Gene Transfer Techniques
Genetic Vectors
/ administration & dosage
Green Fluorescent Proteins
/ genetics
Humans
Induced Pluripotent Stem Cells
/ metabolism
Inteins
Mice
Organoids
/ ultrastructure
Phenotype
Photoreceptor Cells, Vertebrate
/ metabolism
Retina
/ virology
Swine
Trans-Splicing
/ genetics
Journal
Science translational medicine
ISSN: 1946-6242
Titre abrégé: Sci Transl Med
Pays: United States
ID NLM: 101505086
Informations de publication
Date de publication:
15 05 2019
15 05 2019
Historique:
received:
17
09
2018
revised:
21
11
2018
accepted:
04
04
2019
entrez:
17
5
2019
pubmed:
17
5
2019
medline:
27
6
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31092694
pii: 11/492/eaav4523
doi: 10.1126/scitranslmed.aav4523
pmc: PMC6863751
mid: EMS84933
pii:
doi:
Substances chimiques
enhanced green fluorescent protein
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : European Research Council
ID : 670881
Pays : International
Organisme : European Research Council
ID : 694323
Pays : International
Organisme : Telethon
ID : TGM11CB1
Pays : Italy
Informations de copyright
Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.
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