Zhao Yidi, Huang Zhengwei, Zhou Ximeng, Teng Wan, Liu Zehua, Wang Wenping, Tang Shengjia, Liu Ying, Liu Jing, Wang Wenxi, Chai Lingling, Zhang Na, Guo Weilong, Liu Jie, Ni Zhongfu, Sun Qixin, Wang Yanpeng, Zong Yuan
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, China.
Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.
Nat Plants. 2025 Feb;11(2):191-205. doi: 10.1038/s41477-024-01898-3. Epub 2025 Jan 13.
Precise manipulation of genome structural variations holds great potential for plant trait improvement and biological research. Here we present a genome-editing approach, dual prime editing (DualPE), that efficiently facilitates precise deletion, replacement and inversion of large DNA fragments in plants. In our experiments, DualPE enabled the production of specific genomic deletions ranging from ~500 bp to 2 Mb in wheat protoplasts and plants. DualPE was effective in directly replacing wheat genomic fragments of up to 258 kb with desired sequences in the absence of donor DNA. Additionally, DualPE allowed precise DNA inversions of up to 205.4 kb in wheat plants with efficiencies of up to 51.5%. DualPE also successfully edited large DNA fragments in the dicots Nicotiana benthamiana and tomato, with editing efficiencies of up to 72.7%. DualPE thus provides a precise and efficient approach for large DNA sequence and chromosomal engineering, expanding the availability of precision genome-editing tools for crop improvement.
对基因组结构变异进行精确操作在植物性状改良和生物学研究方面具有巨大潜力。在此，我们提出一种基因组编辑方法——双碱基编辑（DualPE），它能有效促进植物中大片段DNA的精确缺失、替换和倒位。在我们的实验中，DualPE能够在小麦原生质体和植株中产生约500 bp至2 Mb的特定基因组缺失。在没有供体DNA的情况下，DualPE能有效地将长达258 kb的小麦基因组片段直接替换为所需序列。此外，DualPE能使小麦植株中长达205.4 kb的DNA精确倒位，效率高达51.5%。DualPE还成功地编辑了双子叶植物本氏烟草和番茄中的大片段DNA，编辑效率高达72.7%。因此，DualPE为大片段DNA序列和染色体工程提供了一种精确且高效的方法，扩大了用于作物改良的精确基因组编辑工具的可用性。
