Efficient and versatile multiplex prime editing in hexaploid wheat
Pei Ni(China Agricultural University), Yidi Zhao(China Agricultural University), Ximeng Zhou(China Agricultural University), Zehua Liu(China Agricultural University), Zhengwei Huang(China Agricultural University), Zhongfu Ni(China Agricultural University), Qixin Sun(China Agricultural University), Yuan Zong(China Agricultural University)
Cited by 106Open Access
Abstract
Prime editing is limited by low efficiency in plants. Here, we develop an upgraded engineered plant prime editor in hexaploid wheat, ePPEplus, by introducing a V223A substitution into reverse transcriptase in the ePPEmax* architecture. ePPEplus enhances the efficiency by an average 33.0-fold and 6.4-fold compared to the original PPE and ePPE, respectively. Importantly, a robust multiplex prime editing platform is established for simultaneous editing of four to ten genes in protoplasts and up to eight genes in regenerated wheat plants at frequencies up to 74.5%, thus expanding the applicability of prime editors for stacking of multiple agronomic traits.