Prime editing: Emerging mechanisms, engineering innovations, and next-generation applications.

Abstract

Prime editing has become a highly programmable and accurate genome-editing platform that can install targeted substitutions, insertions, and deletions without introducing double-strand breaks or requiring a separate donor DNA template. This review summarizes recent developments about prime editing mechanisms, such as knowledge about flap dynamics, repair pathway interactions, and pegRNA architecture, and improvements in engineering, resulting in high-efficiency systems, including PEmax, PE4/5, TWIN-PE, PASTE, and PrimeRoot. Such advances now make prime editing applicable to therapeutic gene correction, agricultural biotechnology, microbial engineering, and functional genomics. However, delivery, chromatin context, mismatch-repair variability, and large-fragment integration remain major barriers to broad application. By comparing prime editing with other genome-editing modalities, this review summarizes its unique advantages and highlights strategic innovations needed for its next stage of development. Together, these developments position prime editing as a highly programmable platform with strong potential to shape the future of precise genome rewriting.