biostudies-literaturePorter JJCystic Fibrosis FoundationNIH HHS14112-14132https://www.ebi.ac.uk/biostudies/studies/S-EPMC11662937biostudies-literatureUnknown52(22)Nonsense suppressor transfer RNAs (tRNAs) or AntiCodon-Edited tRNAs (ACE-tRNAs) have long been envisioned as a therapeutic approach to overcome genetic diseases resulting from the introduction of premature termination codons (PTCs). The ACE-tRNA approach for the rescue of PTCs has been hampered by ineffective delivery through available modalities for gene therapy. Here we have screened a series of ACE-tRNA expression cassette sequence libraries containing >1800 members in an effort to optimize ACE-tRNA function and provide a roadmap for optimization in the future. By optimizing PTC suppression efficiency of ACE-tRNAs, we have decreased the amount of ACE-tRNA required by ∼16-fold for the most common cystic fibrosis-causing PTCs.Nucleic acids researchOptimization of ACE-tRNAs function in translation for suppression of nonsense mutations.PMC11662937PORTER20F0R01 HL153988Porter JJLueck JDKo WSorensen EGfalseOptimization of ACE-tRNAs function in translation for suppression of nonsense mutations.Nonsense suppressor transfer RNAs (tRNAs) or AntiCodon-Edited tRNAs (ACE-tRNAs) have long been envisioned as a therapeutic approach to overcome genetic diseases resulting from the introduction of premature termination codons (PTCs). The ACE-tRNA approach for the rescue of PTCs has been hampered by ineffective delivery through available modalities for gene therapy. Here we have screened a series of ACE-tRNA expression cassette sequence libraries containing >1800 members in an effort to optimize ACE-tRNA function and provide a roadmap for optimization in the future. By optimizing PTC suppression efficiency of ACE-tRNAs, we have decreased the amount of ACE-tRNA required by ∼16-fold for the most common cystic fibrosis-causing PTCs.2024-01-01T00:00:00Z2024 Dec2025-04-04T21:11:33.915Z2025-04-04T21:11:33.915ZS-EPMC116629373967326510.1093/nar/gkae1112