Efficient homologous replacement and deletion of large genomic fragments through template‐jumping prime editing in rice
TL;DR
This study developed template‐jumping prime editing (TJ‐PE), a recently reported PE strategy for large targeted insertion, as an efficient tool for homologous replacement with DNA fragments ranging from dozens to hundreds of base pairs, and using TJ‐PE, replaced genomic fragments of up to 340 bp with homologous fragments of the same length.
Abstract
Homologous replacement of genomic sequences with large DNA fragments (> 100 bp) holds great potential for crop breeding, yet an efficient method to achieve such edits is lacking in plants. Here, in rice, we developed template‐jumping prime editing (TJ‐PE), a recently reported PE strategy for large targeted insertion, as an efficient tool for homologous replacement with DNA fragments ranging from dozens to hundreds of base pairs, and using TJ‐PE, we replaced genomic fragments of up to 340 bp with homologous fragments of the same length. In addition, our TJ‐PE tool also enabled precise deletion of 944‐ to 2024‐bp fragments in rice, with efficiencies of up to 34.6% for c. 2000‐bp precise deletions. Collectively, this study expands the editing scope of PE in rice and establishes TJ‐PE as a generalist tool for precise deletion and replacement of large DNA fragments.