biostudies-literatureTasfaout HWellstone Center at the University of WashingtonUniversity of Washington Center for Translational Muscle ResearchDiabetes Research CenterAssociation Française contre les MyopathiesUS Department of DefenseNational Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of HealthMuscular Dystrophy Associatione189075https://www.ebi.ac.uk/biostudies/studies/S-EPMC12321383biostudies-literatureUnknown135(15)Gene replacement therapies mediated by adeno-associated viral (AAV) vectors represent a promising approach for treating genetic diseases. However, their modest packaging capacity (~4.7 kb) remains an important constraint and significantly limits their application for genetic disorders involving large genes. A prominent example is Duchenne muscular dystrophy (DMD), whose protein product dystrophin is generated from a 11.2 kb segment of the DMD mRNA. Here, we explored methods that enable efficient expression of full-length dystrophin via triple AAV codelivery. This method exploits the protein trans-splicing mechanism mediated by split inteins. We identified a combination of efficient and specific split intein pairs that enabled the reconstitution of full-length dystrophin from 3 dystrophin fThe Journal of clinical investigationExpression of full-length dystrophin reverses muscular dystrophy defects in young and old mdx4cv mice.PMC12321383P30AR074990R01HL128368P30 DK017047P50 AR065139106037224777MD220097Tasfaout HTian RRegnier MMcMillen TSChamberlain JSHalbert CLReyes TRfalseExpression of full-length dystrophin reverses muscular dystrophy defects in young and old mdx4cv mice.Gene replacement therapies mediated by adeno-associated viral (AAV) vectors represent a promising approach for treating genetic diseases. However, their modest packaging capacity (~4.7 kb) remains an important constraint and significantly limits their application for genetic disorders involving large genes. A prominent example is Duchenne muscular dystrophy (DMD), whose protein product dystrophin is generated from a 11.2 kb segment of the DMD mRNA. Here, we explored methods that enable efficient expression of full-length dystrophin via triple AAV codelivery. This method exploits the protein trans-splicing mechanism mediated by split inteins. We identified a combination of efficient and specific split intein pairs that enabled the reconstitution of full-length dystrophin from 3 dystrophin f2025-01-01T00:00:00Z2025 Aug2026-03-31T11:30:53.763Z2025-08-31T03:08:28.761ZS-EPMC123213834049340010.1172/JCI189075