Frontiers Science Center for Molecular Design Breeding (MOE), Key Laboratory of Crop Heterosis and Utilization (MOE), and Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing, 100193, China.
Genome Biol. 2023 Jun 29;24(1):156. doi: 10.1186/s13059-023-02990-1.
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.
在植物中,引发编辑的效率有限。在这里,我们通过在 ePPEmax* 结构中的逆转录酶中引入 V223A 取代,开发了一种在六倍体小麦中经过改良的工程化植物引发编辑器 ePPEplus。与原始的 PPE 和 ePPE 相比,ePPEplus 的效率分别提高了平均 33.0 倍和 6.4 倍。重要的是,建立了一个强大的多路复用引发编辑平台,可在原生质体中同时编辑四个到十个基因,在再生小麦植株中编辑多达八个基因,频率高达 74.5%,从而扩展了引发编辑器在多个农艺性状叠加方面的适用性。