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

ABSTRACT: 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 pr

SUBMITTER: Mu H 

PROVIDER: S-EPMC12069386 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature