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Efficient engineering of human and mouse primary cells using peptide-assisted genome editing.

Zhen ZhangAmy E BaxterDiqiu RenKunhua QinZeyu ChenSierra M CollinsHua HuangChad A KomarPeter F BailerJared B ParkerGerd A BlobelRahul M KohliE John WherryShelley L BergerJunwei Shi
Published in: Nature biotechnology (2023)
Simple, efficient and well-tolerated delivery of CRISPR genome editing systems into primary cells remains a major challenge. Here we describe an engineered Peptide-Assisted Genome Editing (PAGE) CRISPR-Cas system for rapid and robust editing of primary cells with minimal toxicity. The PAGE system requires only a 30-min incubation with a cell-penetrating Cas9 or Cas12a and a cell-penetrating endosomal escape peptide to achieve robust single and multiplex genome editing. Unlike electroporation-based methods, PAGE gene editing has low cellular toxicity and shows no significant transcriptional perturbation. We demonstrate rapid and efficient editing of primary cells, including human and mouse T cells, as well as human hematopoietic progenitor cells, with editing efficiencies upwards of 98%. PAGE provides a broadly generalizable platform for next-generation genome engineering in primary cells.
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