2026-04-13T13:27:39Zhttps://recercat.cat/oai/request
oai:recercat.cat:2117/4143792026-01-25T02:16:46Zcom_2072_1033col_2072_452950
RECERCAT author Truong, Dong-Jiunn Jeffery author Geilenkeuser, Julian author Wendel, Stephanie Victoria author Clemens, Julius author Armbrust, Niklas author Hildegard Binder, Eva Maria author Santl, Tobias Heinrich author Siebenhaar, Annika author Gruber, Christoph author Phlairaharn, Teeradon author Živanić, Milica author Westmeyer, Gil Gregor 2024-03 Prime editing (PE) is a powerful gene-editing technique based on targeted gRNA-templated reverse transcription and integration of the de novo synthesized single-stranded DNA. To circumvent one of the main bottlenecks of the method, the competition of the reverse-transcribed 3' flap with the original 5' flap DNA, we generated an enhanced fluorescence-activated cell sorting reporter cell line to develop an exonuclease-enhanced PE strategy (‘Exo-PE’) composed of an improved PE complex and an aptamer-recruited DNA-exonuclease to remove the 5' original DNA flap. Exo-PE achieved better overall editing efficacy than the reference PE2 strategy for insertions =30¿base pairs in several endogenous loci and cell lines while maintaining the high editing precision of PE2. By enabling the precise incorporation of larger insertions, Exo-PE complements the growing palette of different PE tools and spurs additional refinements of the PE machinery.Peer ReviewedPostprint (published version) http://creativecommons.org/licenses/by/4.0/ Open Access Attribution 4.0 International Àrees temàtiques de la UPC::Enginyeria química::Biotecnologia Genetic engineering Gene regulation Gene targeting Genetic engineering Molecular engineering Enginyeria genètica Exonuclease-enhanced prime editors Article