Prime editing has rapidly emerged as a versatile and precise genome engineering technology capable of introducing targeted substitutions, insertions, and deletions without requiring double-strand breaks or donor DNA templates. Since its initial development, substantial progress has been achieved through iterative innovations in core components, including Cas protein variants, reverse transcriptase engineering, and prime editing guide RNA (pegRNA) design. These advancements have significantly improved editing efficiency, fidelity, and target accessibility across diverse biological systems. In parallel, the development of large-fragment editing platforms such as TwinPE, PASTE, and PrimeRoot has expanded the scope of prime editing from small sequence modifications to kilobase-scale genome rewriting. Furthermore, the integration of artificial intelligence-driven design tools has transformed pegRNA optimisation and system selection, enabling more predictable and efficient editing outcomes. This review provides a comprehensive overview of recent molecular innovations, large-fragment editing strategies, and computational approaches shaping the evolution of prime editing. Collectively, these developments position prime editing as a highly programmable and scalable platform with broad applications in medicine, agriculture, and synthetic biology.

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.