Institute of Cancer Research, Department of Medicine I, Comprehensive Cancer Centre, Medical University of Vienna, Borschkegasse 8a, 1090, Vienna, Austria.
CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
Nat Commun. 2022 Feb 9;13(1):760. doi: 10.1038/s41467-022-28442-1.
Prime editing (PE) is a powerful genome engineering approach that enables the introduction of base substitutions, insertions and deletions into any given genomic locus. However, the efficiency of PE varies widely and depends not only on the genomic region targeted, but also on the genetic background of the edited cell. Here, to determine which cellular factors affect PE efficiency, we carry out a focused genetic screen targeting 32 DNA repair factors, spanning all reported repair pathways. We show that, depending on cell line and type of edit, ablation of mismatch repair (MMR) affords a 2-17 fold increase in PE efficiency, across several human cell lines, types of edits and genomic loci. The accumulation of the key MMR factors MLH1 and MSH2 at PE sites argues for direct involvement of MMR in PE control. Our results shed new light on the mechanism of PE and suggest how its efficiency might be optimised.
碱基编辑（PE）是一种强大的基因组工程方法，可将碱基替换、插入和缺失引入任何给定的基因组位点。然而，PE 的效率差异很大，不仅取决于目标基因组区域，还取决于编辑细胞的遗传背景。在这里，为了确定哪些细胞因素影响 PE 的效率，我们针对 32 种 DNA 修复因子进行了有针对性的遗传筛选，涵盖了所有报道的修复途径。我们发现，根据细胞系和编辑类型，错配修复（MMR）的缺失可使几种人类细胞系、编辑类型和基因组位点的 PE 效率提高 2-17 倍。关键的 MMR 因子 MLH1 和 MSH2 在 PE 位点的积累表明 MMR 直接参与了 PE 的调控。我们的研究结果为 PE 的机制提供了新的见解，并为如何优化其效率提供了线索。
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