文献类型： 外文期刊
作者： Xiong, Youcai 1 ; Su, Yinyu 1 ; He, Ruigao 1 ; Han, Xiaosong 1 ; Li, Sheng 1 ; Liu, Minghuan 1 ; Xi, Xiaoning 1 ; Liu, Zijia 1 ; Wang, Heng 1 ; Xie, Shengsong 1 ; Xu, Xuewen 1 ; Li, Kui 6 ; Zhang, Jifeng 7 ; Xu, Jie 7 ; Li, Xinyun 1 ; Zhao, Shuhong 1 ; Ruan, Jinxue 1 ;
作者机构： 1.Huazhong Agr Univ, Frontiers Sci Ctr Anim Breeding & Sustainable Prod, Wuhan, Peoples R China
2.Huazhong Agr Univ, Key Lab Swine Genet & Breeding Minist Agr & Rural, Breeding & Reprod Minist Educ, Wuhan, Peoples R China
3.Yazhouwan Natl Lab, Sanya, Peoples R China
4.Huazhong Agr Univ, Cooperat Innovat Ctr Sustainable Pig Prod, Wuhan, Peoples R China
5.Huazhong Agr Univ, Hubei Hongshan Lab, Wuhan, Peoples R China
6.Chinese Acad Agr Sci, Agr Genom Inst Shenzhen, Shenzhen, Peoples R China
7.Univ Michigan, Med Sch, Ctr Adv Models Translat Sci & Therapeut, Ann Arbor, MI USA
期刊名称：NATURE COMMUNICATIONS （ 影响因子：15.7； 五年影响因子：17.2 ）
ISSN：
年卷期： 2025 年 16 卷 1 期
页码：
收录情况： SCI
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.
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