Division of Applied Life Science (BK21 Four Program), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Republic of Korea.
Forest Protection Research Centre, Vietnamese Academy of Forest Sciences, Hanoi, Vietnam.
Nat Plants. 2024 Oct;10(10):1502-1513. doi: 10.1038/s41477-024-01786-w. Epub 2024 Sep 6.
Prime editing (PE) enables almost all types of precise genome editing in animals and plants. It has been successfully adapted to edit several plants with variable efficiency and versatility. However, this technique is inefficient for dicots for unknown reasons. Here, using new combinations of PE components, including an RNA chaperone and altered engineered prime editing guide RNAs driven by a PolII-PolIII composite promoter and a viral replicon system, we obtained up to 9.7% of the desired PE efficiency at the callus stage as assessed by targeted deep sequencing. Subsequently, we identified that up to 38.2% of transformants contained desired PE alleles in tomatoes and Arabidopsis, marking successful heritable PE transmission. Our PE tools also showed high accuracy, specificity and multiplexing capability, which unlocked the potential for practical PE applications in dicots, paving the way for transformative advancements in plant sciences.
先导编辑(PE)可实现动植物中几乎所有类型的精确基因组编辑。它已成功应用于编辑几种具有不同效率和多功能性的植物。然而,由于未知原因,该技术在双子叶植物中效率低下。在这里,我们使用包括 RNA 伴侣和经过工程改造的先导编辑引导 RNA 的新组合,这些 RNA 由 PolII-PolIII 复合启动子和病毒复制子系统驱动,在愈伤组织阶段通过靶向深度测序评估,我们获得了高达 9.7%的所需 PE 效率。随后,我们在番茄和拟南芥中鉴定出高达 38.2%的转化体含有所需的 PE 等位基因,标志着可遗传的 PE 传递成功。我们的 PE 工具还表现出高精度、特异性和多重编辑能力,为双子叶植物中实用的 PE 应用开辟了道路,为植物科学的变革性进步铺平了道路。