先导编辑技术（prime editing）在双子叶植物中的优化

先导编辑技术（prime editing）作为一种新型基因编辑技术，无需供体DNA、不引起DNA双链断裂、能同时实现小片段DNA序列的增删和碱基12种类型的转换，被誉为“改变基因编辑游戏规则的工具”。然而作为新兴的基因编辑工具，其编辑效率较低，亟待优化。在植物中实施和优化先导编辑，对于植物基因编辑工具开发更有针对性。本项目前期在双子叶模式植物——本氏烟草中成功实施了先导编辑。后续将以建立快速检测先导编辑效率的报告系统为基础，筛选适合于植物的高效逆转录组件。本项目将采取多策略的逆转录组件优化方案，采用最优转化和培养方法，最终整合各优化措施以加合效应提高双子叶植物先导编辑效率。高效的双子叶植物先导编辑系统将作为有利的工具服务于作物育种（产生、聚合优良性状）和植物合成生物学（植物天然产物的合成通路解析），创造更大的科学价值和经济价值。

Prime editing is a new breakthrough of genome editing tool without donor DNA or double-strand breaks, allowing introduction of all mutation types (insertions, deletions, and all 12 types of base-to-base conversions). However, quite low editing efficiency reported by researchers when prime editing was applied in many species. Optimizing prime editor in plants is more targeted to develop useful plant genome editing tools. Having successfully applied prime editing in Nicotiana benthamiana, we focus on establishing a fast and sensitive detecting system to evaluate editing efficiency by prime editing recovering LUC activity. With this detecting system, we plan to replace the reverse transcriptase and increase the copy number of reverse transcriptase to make prime editor suitable for plant prime editing. Multiple optimizations of prime editor and more effective transformation method would likely make prime editing work better in plants by additive effect. This project aims to enhance the editing efficiency of plant prime editing which would be a powerful genome editing tool in crop breeding and plant synthetic biology.