Nonsense mutations are the underlying cause of approximately 11% of all inherited genetic diseases¹. Nonsense mutations convert a sense codon that is decoded by tRNA into a premature termination codon (PTC), resulting in an abrupt termination of translation. One strategy to suppress nonsense mutations is to use natural tRNAs with altered anticodons to base-pair to the newly emerged PTC and promote translation^2,3,4,5,6,7. However, tRNA-based gene therapy has not yielded an optimal combination of clinical efficacy and safety and there is presently no treatment for individuals with nonsense mutations. Here we introduce a strategy based on altering native tRNAs into efficient suppressor tRNAs (sup-tRNAs) by individually fine-tuning their sequence to the physico-chemical properties of the amino acid that they carry. Intravenous and intratracheal lipid nanoparticle (LNP) administration of sup-tRNA in mice restored the production of functional proteins with nonsense mutations. LNP–sup-tRNA formulations caused no discernible readthrough at endogenous native stop codons, as determined by ribosome profiling. At clinically important PTCs in the cystic fibrosis transmembrane conductance regulator gene (CFTR), the sup-tRNAs re-established expression and function in cell systems and patient-derived nasal epithelia and restored airway volume homeostasis. These results provide a framework for the development of tRNA-based therapies with a high molecular safety profile and high efficacy in targeted PTC suppression.

无义突变是大约 11% 的遗传性遗传疾病的根本原因¹。无义突变将 tRNA 解码的有义密码子转化为过早终止密码子 (PTC)，导致翻译突然终止。抑制无义突变的一种策略是使用具有改变的反密码子的天然 tRNA 与新出现的 PTC 进行碱基配对并促进翻译^2,3,4,5,6,7。然而，基于 tRNA 的基因治疗尚未产生临床疗效和安全性的最佳组合，并且目前还没有针对具有无义突变的个体的治疗方法。在这里，我们介绍了一种基于将天然 tRNA 改变为有效抑制 tRNA (sup-tRNA) 的策略，通过单独微调它们的序列以适应它们携带的氨基酸的物理化学特性。对小鼠进行静脉内和气管内脂质纳米颗粒（LNP）注射sup-tRNA可恢复具有无义突变的功能蛋白的产生。根据核糖体分析确定，LNP-sup-tRNA 制剂在内源性天然终止密码子处没有引起明显的通读。在囊性纤维化跨膜电导调节基因 ( CFTR ) 的临床重要 PTC 中，sup-tRNA 重新建立了细胞系统和患者来源的鼻上皮细胞中的表达和功能，并恢复了气道容量稳态。这些结果为开发基于 tRNA 的疗法提供了框架，该疗法具有高分子安全性和靶向 PTC 抑制的高效性。