IJMS, Vol. 26, Pages 7943: Prime Editing Modification with FEN1 Improves F508del Variant Editing in the CFTR Gene in Airway Basal Cells

International Journal of Molecular Sciences doi: 10.3390/ijms26167943

Authors:
Olga V. Volodina
Anna G. Demchenko
Arina A. Anuchina
Oxana P. Ryzhkova
Valeriia A. Kovalskaya
Ekaterina V. Kondrateva
Ekaterina V. Artemova
Vyacheslav Y. Tabakov
Maxim A. Ignatov
Natalia Y. Vorobyeva
Andreyan N. Osipov
Alexander V. Lavrov
Svetlana A. Smirnikhina

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 CFTR 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.

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Olga V. Volodina www.mdpi.com