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

Liu, Bin; Dong, Xiaolong; Zheng, Chunwei; Keener, David; Chen, Zexiang; Cheng, Haoyang; Watts, Jonathan K; Xue, Wen et al. · Nat Biotechnol · 2024

basic_science · Level V

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Abstract

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).

Medical subject headings