南湖新闻网讯（通讯员 阮进学 熊友才）近日，我校动物遗传育种团队研究成果以“EXPERT expands prime editing efficiency and range of large fragment edits”为题在Nature Communications发表。研究开发了新型基因编辑工具EXPERT，创新性地在pegRNA nick所在DNA链上游引入额外切口，并对pegRNA末端进行优化改造，同时引入突变以抑制游离片段对新链合成的干扰，大幅度拓宽了PE编辑范围，在多个物种中实现了大片段精准高效编辑，为生物育种、人类疾病治疗等领域提供了新技术和工具。

▲EXPERT技术原理模式图

Prime Editing（PE）作为一种精准基因编辑技术，相较于BE（Base Editing）技术具有碱基替换灵活、编辑种类更多、更精准等优点，但也存在编辑范围受限、长片段编辑效率较低、在猪等动物中编辑效率低等不足，这在一定程度上制约了PE技术的发展和应用。针对这些关键问题，我校动物遗传育种团队成功研发出EXPERT基因编辑技术，该技术创新性地通过在pegRNA nick所在DNA链上游引入额外切口、对pegRNA末端改造以及引入突变抑制游离片段对新链合成干扰等手段进行优化，在多个物种中实现了大片段高效精准编辑。

EXPERT相比传统PE系统具有三个主要优势：一是更广泛的编辑范围，EXPERT能够同时编辑pegRNA nick的上游和下游区域，可实现100bp以上片段高效精准编辑，极大地拓宽了传统PE编辑范围。二是更高的编辑效率，与PE2系统相比，EXPERT在长片段（插入、删除、替换）编辑效率最高提升达122倍，同时保持较低的非预期indel比率，实现了大片段高效精准编辑。三是在猪等动物中具有更高的编辑效率，传统的PE系统在猪中活性不高，EXPERT在猪、鼠、人中都具有很高的编辑效率，显示出良好跨物种基因编辑能力，为生物育种和人类疾病治疗提供了新技术和工具。

华中农业大学动科动医学院博士毕业生熊友才、博士研究生苏寅钰和何瑞高为该论文共同第一作者，赵书红教授、阮进学副研究员和李新云教授为共同通讯作者。该研究得到了国家重点研发项目、国家自然科学基金创新群体项目等项目资助。

审核人：李新云

【英文摘要】

Prime editing systems (PEs) hold great promise in modern biotechnology. However, their editing range is limited as PEs can only modify the downstream sequences of the pegRNA nick. Here, we report the development of the extended prime editor system (EXPERT) to overcome this limitation by using an extended pegRNA (ext-pegRNA) with modified 3’ extension, and an additional sgRNA (ups-sgRNA) targeting the upstream region of the ext-pegRNA. We demonstrate that EXPERT can efficiently perform editing on both sides of the ext-pegRNA nick, a task that is unattainable by canonical PEs. EXPERT exhibits prominent capacity in replacing sequences up to 88 base pairs and inserting sequences up to 100 base pairs within the upstream region of the ext-pegRNA nick. Compared to canonical PEs such as PE2, the utilization of the EXPERT strategy significantly enhances the editing efficiency for large fragment edits with an average improvement of 3.12-fold, up to 122.1 times higher. Safety wise, the use of ups-sgRNA does not increase the rates of undesirable insertions and deletions (indels), as the two nicks are on the same strand. Moreover, we do not observe increased off-target editing rates genome-wide. Our work introduces EXPERT as a PE tool with significant potential in life sciences.

论文链接：https://www.nature.com/articles/s41467-025-56734-9?utm_source=rct_congratemailt&utm_medium=email&utm_campaign=oa_20250213&utm_content=10.1038/s41467-025-56734-9