Volodina Olga V, Demchenko Anna G, Anuchina Arina A, Ryzhkova Oxana P, Kovalskaya Valeriia A, Kondrateva Ekaterina V, Artemova Ekaterina V, Tabakov Vyacheslav Y, Ignatov Maxim A, Vorobyeva Natalia Y, Osipov Andreyan N, Lavrov Alexander V, Smirnikhina Svetlana A
Laboratory of Genome Editing, Research Centre for Medical Genetics, 115522 Moscow, Russia.
Laboratory for Structural Analysis and Engineering of Membrane Systems, The Research Center for Molecular Mechanisms of Aging and Age-Related Diseases, Moscow Institute of Physics and Technology, 141701 Moscow, Russia.
Int J Mol Sci. 2025 Aug 18;26(16):7943. doi: 10.3390/ijms26167943.
Prime editing is a promising approach for correcting pathogenic variants, but its efficiency remains variable across genomic contexts. Here, we systematically evaluated 12 modifications of the PEmax system for correcting the F508del pathogenic variant that caused cystic fibrosis in patient-derived airway basal cells. We chose EXO1 and FEN1 nucleases to improve the original system. While all tested variants showed comparatively low efficiency in this AT-rich genomic region, 4-FEN modification demonstrated significantly improved editing rates (up to 2.13 fold) compared to standard PEmax. Our results highlight two key findings: first, the persistent challenge of AT-rich target sequence correction even with optimized editors, and second, the performance of 4-FEN suggests its potential value for other genomic targets.
碱基编辑是一种很有前景的纠正致病变体的方法，但其效率在不同的基因组背景下仍存在差异。在这里，我们系统地评估了PEmax系统的12种修饰，以纠正导致患者来源的气道基底细胞囊性纤维化的F508del致病变体。我们选择了EXO1和FEN1核酸酶来改进原始系统。虽然所有测试变体在这个富含AT的基因组区域都表现出相对较低的效率，但与标准PEmax相比，4-FEN修饰的编辑率显著提高（高达2.13倍）。我们的结果突出了两个关键发现：第一，即使使用优化的编辑器，富含AT的靶序列校正仍然存在持续挑战；第二，4-FEN的性能表明其对其他基因组靶点具有潜在价值。
