Duchenne muscular dystrophy (DMD) is a fatal X-chromosome-linked genetic disease caused by dystrophin gene mutations, including nonsense mutations.1 Nonsense mutations are caused by the introduction of premature termination codons, which prevent translation of full-length proteins. Read-through therapies show potential for addressing DMD's genetic basis; however, issues such as non-specific amino acid insertions, gene-editing delivery challenges, and clinical safety concerns have limited their progress.2,3 To address nonsense mutations, nonsense suppressor transfer RNAs (sup-tRNAs) have been proposed as a genetic therapy approach.3, 4, 5 In this study, we propose a new MyoAAV-delivered suppressor tRNA (sup-tRNA) strategy to restore dystrophin expression. Our approach specifically targets nonsense mutations in mdx mice and patient-derived myoblasts and cardiomyocytes, significantly increasing dystrophin levels, especially in the heart (up to 61.43% when combined with CC-90009). This combination alleviates dystrophic symptoms and improves read-through efficiency, likely by reducing translation termination factor activity. These findings highlight the potential of sup-tRNA in DMD and other nonsense mutation-related diseases.