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Article

High-yield genome engineering in primary cells using a hybrid ssDNA repair template and small-molecule cocktails.

Nature biotechnology - 1 Apr 2023

Shy Brian R, Vykunta Vivasvan S, Ha Alvin, Talbot Alexis, Roth Theodore L, Nguyen David N, Pfeifer Wolfgang G, Chen Yan Yi, Blaeschke Franziska, Shifrut Eric, Vedova Shane, Mamedov Murad R, Chung Jing-Yi Jing, Li Hong, Yu Ruby, Wu David, Wolf Jeffrey, Martin Thomas G, Castro Carlos E, Ye Lumeng, Esensten Jonathan H, Eyquem Justin, Marson Alexander

Abstract excerpt

Enhancing CRISPR-mediated site-specific transgene insertion efficiency by homology-directed repair (HDR) using high concentrations of double-stranded DNA (dsDNA) with Cas9 target sequences (CTSs) can be toxic to primary cells. Here, we develop single-stranded DNA (ssDNA) HDR templates (HDRTs) incorporating CTSs with reduced toxicity that boost knock-in efficiency and yield by an average of around two- to...

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Methodology Data sources Editorial policy Privacy About Contact Helena Bioinformatics

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Source

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Identifiers

PMID: 36008610
DOI: 10.1038/s41587-022-01418-8
PMCID: PMC10065198

Provenance

PubMed metadata and Literature Corpus relationships. Corpus snapshot updated 7 Sept 2026. Manifest publication-graph-cohort-2026-09-07.1.

Source receipt: pubmed, retrieved 7 Sept 2026.

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