Programmed RNA editing presents an attractive therapeutic strategy for genetic disease. In this study, we developed bacterial deaminase-enabled recoding of RNA (DECOR), which employs an evolved Escherichia coli transfer RNA adenosine deaminase, TadA8e, to deposit adenosine-to-inosine editing to CRISPR-specified sites in the human transcriptome. DECOR functions in a variety of cell types, including human lung fibroblasts, and delivers on-target activity similar to ADAR-overexpressing RNA-editing platforms with 88% lower off-target effects. High-fidelity DECOR further reduces off-target effects to basal level. We demonstrate the clinical potential of DECOR by targeting Van der Woude syndrome-causing interferon regulatory factor 6 (IRF6) insufficiency. DECOR-mediated RNA editing removes a pathogenic upstream open reading frame (uORF) from the 5′ untranslated region of IRF6 and rescues primary ORF expression from 12.3% to 36.5%, relative to healthy transcripts. DECOR expands the current portfolio of effector proteins and opens new territory in programmed RNA editing.

程序化 RNA 编辑为遗传病提供了一种有吸引力的治疗策略。在这项研究中，我们开发了细菌脱氨酶启用的 RNA 重编码 （DECOR），它采用进化的大肠杆菌转移 RNA 腺苷脱氨酶 TadA8e，将腺苷-肌苷编辑沉积到人类转录组中的 CRISPR 指定位点。DECOR 在多种细胞类型中发挥作用，包括人肺成纤维细胞，并提供类似于 ADAR 过表达 RNA 编辑平台的靶向活性，脱靶效应降低 88%。高保真 DECOR 进一步将脱靶效应降低到基础水平。我们通过靶向引起 Van der Woude 综合征的干扰素调节因子 6 （IRF6） 不足来证明 DECOR 的临床潜力。DEOR 介导的 RNA 编辑从 IRF6 的 5' 非翻译区去除致病性上游开放阅读框 （uORF），相对于健康转录本，将初级 ORF 表达从 12.3% 恢复到 36.5%。DECOR 扩展了当前的效应蛋白产品组合，并在程序化 RNA 编辑方面开辟了新领域。