德国慕尼黑工业大学Gil Gregor Westmeyer团队开发出外切酶增强的先导编辑器。相关论文于2024年2月1日在线发表于国际学术期刊《自然—方法学》。

研究人员表示，先导编辑（PE）是一种功能强大的基因编辑技术，它以靶向gRNA为模板进行反转录并整合从头合成的单链DNA。

为了规避该方法的主要瓶颈之一——反转录的3′翼与原始5′翼DNA的竞争，研究人员生成了一种增强型荧光激活细胞分选报告细胞系，开发了一种外切酶增强型PE策略（"Exo-PE"），该策略由一种改进的PE复合物和一种诱导DNA外切酶组成，以去除5′原始DNA翼。与参考PE2策略相比，Exo-PE在多个内源基因位点和细胞系中对≥30个碱基对的插入实现了更好的整体编辑效果，同时保持了PE2的高编辑精度。Exo-PE能够精确地整合更大的插入片段，是对日益增多的各种PE工具的补充，并促进PE机器的进一步完善。

附：英文原文

Title: Exonuclease-enhanced prime editors

Author: Truong, Dong-Jiunn Jeffery, Geilenkeuser, Julian, Wendel, Stephanie Victoria, Wilming, Julius Clemens Heinrich, Armbrust, Niklas, Binder, Eva Maria Hildegard, Santl, Tobias Heinrich, Siebenhaar, Annika, Gruber, Christoph, Phlairaharn, Teeradon, ivani, Milica, Westmeyer, Gil Gregor

Issue&Volume: 2024-02-01

Abstract: Prime editing (PE) is a powerful gene-editing technique based on targeted gRNA-templated reverse transcription and integration of the de novo synthesized single-stranded DNA. To circumvent one of the main bottlenecks of the method, the competition of the reverse-transcribed 3′ flap with the original 5′ flap DNA, we generated an enhanced fluorescence-activated cell sorting reporter cell line to develop an exonuclease-enhanced PE strategy (‘Exo-PE’) composed of an improved PE complex and an aptamer-recruited DNA-exonuclease to remove the 5′ original DNA flap. Exo-PE achieved better overall editing efficacy than the reference PE2 strategy for insertions ≥30base pairs in several endogenous loci and cell lines while maintaining the high editing precision of PE2. By enabling the precise incorporation of larger insertions, Exo-PE complements the growing palette of different PE tools and spurs additional refinements of the PE machinery.

DOI: 10.1038/s41592-023-02162-w

Source: https://www.nature.com/articles/s41592-023-02162-w