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• W3176362204 abstract "ABSTRACT Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. Currently, the delivery of PE components to mammalian cell lines requires multiple plasmid vectors. To overcome this limitation, we generated all-in-one prime editing (PEA1) constructs that carry all the components required for PE, along with a selection marker. We tested these constructs (with selection) in HEK293T, K562, HeLa and mouse embryonic stem (ES) cells. We discovered that PE efficiency in HEK293T cells was much higher than previously observed, reaching up to 95% (mean 67%). The efficiency in K562 and HeLa cells, however, remained low. To improve PE efficiency in K562 and HeLa, we generated a nuclease prime editor and tested this system in these cell lines as well as mouse ES cells. PE-nuclease greatly increased prime editing initiation, however, installation of the intended edits was often accompanied by extra insertions derived from the repair template. Finally, we show that zygotic injection of the nuclease prime editor can generate correct modifications in mouse fetuses with up to 100% efficiency. In summary, PE-nuclease and the PEA1 plasmids provide new tools to generate intended edits with high efficiency." @default.
• W3176362204 created "2021-07-05" @default.
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• W3176362204 date "2021-07-02" @default.
• W3176362204 modified "2023-10-09" @default.
• W3176362204 title "Optimized nickase- and nuclease-based prime editing in human and mouse cells" @default.
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• W3176362204 doi "https://doi.org/10.1101/2021.07.01.450810" @default.
• W3176362204 hasPublicationYear "2021" @default.
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