Prime editing is a recently reported genome editing tool using a nickase-cas9 fused to a reverse transcriptase that directly synthesizes the desired edit at the target site. Here, we explore the use of prime editing in human organoids. Common TP53 mutations can be correctly modeled in human adult stem cell-derived colonic organoids with efficiencies up to 25% and up to 97% in hepatocyte organoids. Next, we functionally repaired the cystic fibrosis CFTR-F508del mutation and compared prime editing to CRISPR/Cas9-mediated homology-directed repair and adenine base editing on the CFTR-R785* mutation. Whole-genome sequencing of prime editing-repaired organoids revealed no detectable off-target effects. Despite encountering varying editing efficiencies and undesired mutations at the target site, these results underline the broad applicability of prime editing for modeling oncogenic mutations and showcase the potential clinical application of this technique, pending further optimization.

中文翻译：

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评估基于 CRISPR 的癌症建模和类器官 CFTR 修复的 Prime 编辑。


Prime编辑是最近报道的一种基因组编辑工具，使用与逆转录酶融合的切口酶-cas9，直接在目标位点合成所需的编辑。在这里，我们探索了主要编辑在人类类器官中的应用。常见的 TP53 突变可以在人类干细胞衍生的结肠类器官中正确建模，效率高达 25%，在肝细胞类器官中高达 97%。接下来，我们对囊性纤维化 CFTR-F508del 突变进行功能性修复，并将 Prime 编辑与 CRISPR/Cas9 介导的同源定向修复和 CFTR-R785* 突变上的腺嘌呤碱基编辑进行比较。对主要编辑修复的类器官进行全基因组测序，未发现可检测到的脱靶效应。尽管在目标位点遇到了不同的编辑效率和不需要的突变，但这些结果强调了初等编辑在致癌突变建模中的广泛适用性，并展示了该技术的潜在临床应用，有待进一步优化。