Jeong Yeong Yeop, Hong Cheljong, Han Jun Hee, Bae Sangsu, Seo Pil Joon
Plant Genomics and Breeding Institute, Seoul National University, Seoul 08826; Department of Chemistry, Seoul National University, Seoul 08826; Department of Biological Sciences, Sungkyunkwan University, Suwon 16419, Korea.
Department of Chemistry, Seoul National University, Seoul 08826, Korea.
BMB Rep. 2025 Feb;58(2):70-74. doi: 10.5483/BMBRep.2024-0137.
Prime editing is widely used in many organisms to introduce site-specific sequence modifications such as base substitutions, insertions, and deletions in genomic DNA without generating double-strand breaks. Despite wide-ranging applications of prime editing, prime editors (PEs) have low editing efficiencies, especially in dicot plants. Therefore, PEs are barely used for genome engineering in dicot plant species. Here, based on previous approaches used to improve prime editing efficiency, we generated different combinations of PE components and prime editing guide RNAs (pegRNAs) and examined their prime editing efficiencies in Arabidopsis thaliana protoplasts as a dicot model system. We found that v4e2, in which PE was fused to viral nucleocapsid (NC) protein, RNase Hdeleted M-MLV RT, and a dominant negative version of human mutL homolog 1 (hMLH1dn), showed the highest prime editing efficiency in Arabidopsis protoplasts when it was co-transfected with dual enhanced pegRNA. Our results suggest that the v4e2 PE system could be used for efficient prime editing in dicot plants. [BMB Reports 2025; 58(2): 70-74].
引导编辑被广泛应用于许多生物体中，以在基因组DNA中引入位点特异性序列修饰，如碱基替换、插入和缺失，而不会产生双链断裂。尽管引导编辑有广泛的应用，但引导编辑器（PEs）的编辑效率较低，尤其是在双子叶植物中。因此，PEs几乎未用于双子叶植物物种的基因组工程。在此，基于先前用于提高引导编辑效率的方法，我们生成了PE组件和引导编辑向导RNA（pegRNAs）的不同组合，并在作为双子叶模型系统的拟南芥原生质体中检测了它们的引导编辑效率。我们发现，当v4e2与双增强pegRNA共转染时，其在拟南芥原生质体中显示出最高的引导编辑效率，v4e2是将PE与病毒核衣壳（NC）蛋白、核糖核酸酶H缺失的莫洛尼鼠白血病病毒逆转录酶（M-MLV RT）以及人MutL同源蛋白1的显性负性版本（hMLH1dn）融合而成。我们的结果表明，v4e2 PE系统可用于双子叶植物的高效引导编辑。[《BMB报告》2025年；58(2): 70 - 74]
