植物基因组编辑新工具——引导编辑技术

Plant prime editing technique: a new genome editing tool for plants

基于CRISPR/Cas9系统的引导编辑(prime editing, PE)技术作为一种新兴的基因组编辑技术,能在不产生双链断裂的情况下实现所有12种单碱基替换和小片段DNA的缺失或插入。引导编辑技术已经在多种植物中成功应用并将在植物精准育种中发挥重要作用。虽然植物引导编辑(plant prime editing, PPE)技术极大地扩展了植物基因组精准编辑的范围和能力,但是其编辑效率仍需进一步改善才能推动其在植物基础研究及育种中的广泛应用。文中介绍了植物引导编辑的开发历程、组成结构、优点和局限性,并重点介绍了植物引导编辑效率优化的进展,包括Tm值指导的PBS序列设计、RT模板长度、双pegRNA策略、PlantPegDesigner网站的开发和PPE效应蛋白优化策略。最后,对植物引导编辑技术的未来发展和应用提出了展望。

The CRISPR/Cas9 based prime editing(PE) technique enables all 12 types of base substitutions and precise small DNA deletions or insertions without generating DNA double-strand breaks. Prime editing has been successfully applied in plants and plays important roles in plant precision breeding. Although plant prime editing(PPE) can substantially expand the scope and capabilities of precise genome editing in plants, its editing efficiency still needs to be further improved. Here, we review the development of PPE technique, and introduce structural composition, advantages and limitations of PPE. Strategies to improve the PPE editing efficiency, including the Tm-directed PBS length design, the RT template length, the dual-pegRNA strategy, the PlantPegDesigner website, and the strategies for optimizing the target proteins of PPE, were highlighted. Finally, the prospects of future development and application of PPE were discussed.