*Correspondence: zongyuan@cau.edu.cn

As wheat is a pivotal staple crop globally, maintaining and enhancing its yield is a crucial task. However, the hexaploid nature and functional gene redundancy of wheat pose challenges in effectively and precisely modifying genes for critical agronomic trait improvement. The advent of genome editing technologies has ushered in new hopes for crop improvement.

Nevertheless, CRISPR/Cas9 currently primarily focuses on knocking out wheat genes, leading to functional deficiencies. To address this, we have previously established a precise editing system for single-base editing and predictable multi-nucleotide deletion in wheat, yet it still falls short of meeting the diverse needs in wheat gene function research and molecular breeding.

The Prime Editor (PE) system enables the substitution of any base, as well as the insertion and deletion of small fragments in plant genomes.

However, its editing efficiency in polyploid plants like wheat is extremely low, rendering it nearly unusable. Therefore, our team has comprehensively redesigned the PE system, encompassing the pegRNA, reverse transcriptase, and fusion protein construction, resulting in a highly efficient reengineered plant Prime Editor.

This advancement significantly improved the editing efficiency of the Prime Editor system in hexaploid wheat (by an average of 33-fold) and established an efficient, precise, and stable multi-gene Prime Editing system. We have achieved precise editing of eight genes (21 gene loci) simultaneously in wheat, broadening the applicability of the Prime Editor system in plant genome editing.

This research marks the first report of utilizing the Prime Editor system to obtain wheat mutants, providing significant insights and technical support for Prime Editing and trait stacking in wheat and other polyploid plants.

Furthermore, we have preliminarily achieved cross-scale editing of the common wheat genome, encompassing diverse chromosomal rearrangements such as deletions, duplications, and fusions, using genome editing technology. This offers diverse technical support for various agricultural applications, effectively advancing the process of crop molecular design breeding.