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Systematic optimization of prime editing for enhanced efficiency and versatility in genome engineering across diverse cell types

文献类型：期刊论文

作者Mu, Huiling3,4; Liu, Yeyi2,3; Chi, Yijia2,3; Wang, Fei3; Meng, Shuting1,3; Zhang, Yi3,4; Wang, Xunting3,4; Zhao, Dongxin2,3,4
刊名FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
出版日期2025-04-29
卷号13页码:13
关键词prime editing piggyBac transposon system pluripotent stem cells sustained expression genome engineering
ISSN号2296-634X
DOI10.3389/fcell.2025.1589034
通讯作者Zhao, Dongxin(zhaodongxin@simm.ac.cn)
英文摘要Prime editing offers remarkable versatility in genome editing, but its efficiency remains a major bottleneck. While continuous optimization of the prime editing enzymes and guide RNAs (pegRNAs) has improved editing outcomes, the method of delivery also plays a crucial role in overall performance. To maximize prime editing efficiency, we implemented a series of systematic optimizations, achieving up to 80% editing efficiency across multiple loci and cell lines. Beyond integrating the latest advancements in prime editing, our approach combined stable genomic integration of prime editors via the piggyBac transposon system, selection of integrated single clones, the use of an enhanced promoter, and lentiviral delivery of pegRNAs, ensuring robust, ubiquitous, and sustained expression of both prime editors and pegRNAs. To further assess its efficacy in challenging cell types, we validated our optimized system in human pluripotent stem cells (hPSCs) in both primed and na & iuml;ve states, achieving substantial editing efficiencies of up to 50%. Collectively, our optimized prime editing strategy provides a highly efficient and versatile framework for genome engineering in vitro, serving as a roadmap for refining prime editing technologies and expanding their applications in genetic research and therapeutic development.
WOS关键词BACULOVIRUS GENOME ; GENE-TRANSFER ; PIGGYBAC ; TRANSPOSASES ; INTEGRATION ; NAIVE ; BASE ; DNA
资助项目National Natural Science Foundation of China[32171447] ; National Natural Science Foundation of China[82373972] ; Strategic Priority Research Program of the Chinese Academy of Sciences[XDB0830300]
WOS研究方向Cell Biology ; Developmental Biology
语种英语
WOS记录号WOS:001485683200001
出版者FRONTIERS MEDIA SA
源URL[http://119.78.100.183/handle/2S10ELR8/317908]  
专题中国科学院上海药物研究所
通讯作者Zhao, Dongxin
作者单位1.Henan Univ, Sch Pharm, Kaifeng, Peoples R China
2.Univ Chinese Acad Sci, Sch Pharm, Beijing, Peoples R China
3.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai, Peoples R China
4.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing, Peoples R China
推荐引用方式
GB/T 7714		 Mu, Huiling,Liu, Yeyi,Chi, Yijia,et al. Systematic optimization of prime editing for enhanced efficiency and versatility in genome engineering across diverse cell types[J]. FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY,2025,13:13.
APA Mu, Huiling.,Liu, Yeyi.,Chi, Yijia.,Wang, Fei.,Meng, Shuting.,...&Zhao, Dongxin.(2025).Systematic optimization of prime editing for enhanced efficiency and versatility in genome engineering across diverse cell types.FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY,13,13.
MLA Mu, Huiling,et al."Systematic optimization of prime editing for enhanced efficiency and versatility in genome engineering across diverse cell types".FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY 13(2025):13.

入库方式： OAI收割

来源：上海药物研究所

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