Targeted genome editing with a DNA-dependent DNA polymerase and exogenous DNA-containing templates.

Reverse transcriptases, used in prime editing systems, exhibit lower fidelity, processivity and dNTP affinity than many DNA-dependent DNA polymerases. We report that a DNA-dependent DNA polymerase (phi29), untethered from Cas9, enables editing from a synthetic, end-stabilized DNA-containing template at up to 60% efficiency in human cells. Compared to prime editing, DNA polymerase editing avoids autoinhibitory intramolecular base pairing of the template, facilitates template synthesis and supports larger insertions (>100 nucleotides). © 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.

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Bin Liu, Xiaolong Dong, Chunwei Zheng, David Keener, Zexiang Chen, Haoyang Cheng, Jonathan K Watts, Wen Xue, Erik J Sontheimer. Targeted genome editing with a DNA-dependent DNA polymerase and exogenous DNA-containing templates. Nature biotechnology. 2024 Jul;42(7):1039-1045

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PMID: 37709915

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