基因组错误最少的工程化碱基编辑器。

Engineered prime editors with minimal genomic errors.

作者信息

Chauhan Vikash P, Sharp Phillip A, Langer Robert

机构信息

Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.

Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.

出版信息

Nature. 2025 Sep 17. doi: 10.1038/s41586-025-09537-3.

DOI:10.1038/s41586-025-09537-3

PMID:40963020

Abstract

Prime editors make programmed genome modifications by writing new sequences into extensions of nicked DNA 3' ends. These edited 3' new strands must displace competing 5' strands to install edits, yet a bias towards retaining the competing 5' strands hinders efficiency and can cause indel errors. Here we discover that nicked end degradation, consistent with competing 5' strand destabilization, can be promoted by Cas9-nickase mutations that relax nick positioning. We exploit this mechanism to engineer efficient prime editors with strikingly low indel errors. Combining this error-suppressing strategy with the latest efficiency-boosting architecture, we design a next-generation prime editor (vPE). Compared with previous editors, vPE features comparable efficiency yet up to 60-fold lower indel errors, enabling edit:indel ratios as high as 543:1.

摘要

碱基编辑器通过将新序列写入切口DNA 3'端的延伸部分来进行可编程的基因组修饰。这些编辑后的3'新链必须取代竞争的5'链以安装编辑，但倾向于保留竞争的5'链会阻碍效率并可能导致插入缺失错误。在这里，我们发现与竞争的5'链不稳定一致的切口末端降解可以通过放宽切口定位的Cas9切口酶突变来促进。我们利用这一机制设计了具有极低插入缺失错误的高效碱基编辑器。将这种错误抑制策略与最新的效率提升架构相结合，我们设计了下一代碱基编辑器（vPE）。与以前的编辑器相比，vPE具有相当的效率，但插入缺失错误降低了多达60倍，编辑与插入缺失的比率高达543:1。