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Updated: Jul 16, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
Targeted genome editing with a DNA-dependent DNA polymerase and exogenous DNA-containing templates
Bin Liu1, Xiaolong Dong1,2, Chunwei Zheng1
1RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, MA, USA.
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).
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