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Error-free and efficient prime editing in absence of maternal Polθ

Kroll, F.; Serafini, M.; de Barbarin, L.; Alvarez Vargas, J. R.; Dang, J. T.-M.; Rosello, M.; As, M.; De Cian, A.; Concordet, J.-P.; Giovannangeli, C.; Del Bene, F.

2026-02-04 genetics
10.64898/2026.02.02.703001 bioRxiv
Show abstract

Prime editing was developed to make small edits while minimising the unwanted mutations associated with genome editing based on double-strand breaks. In zebrafish embryos, however, prime editing is notably error-prone, typically yielding more unwanted mutations than precise edits. We show that virtually every unwanted mutation is generated by DNA polymerase {theta} (Pol{theta})-dependent microhomology-mediated end-joining. Loss of maternally-deposited Pol{theta} abolishes unwanted mutations and can increase edit rates above 50%, resulting in error-free and efficient prime editing.

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