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

Peptide-assisted genome editing enables efficient single and multiplex editing in hematopoietic cells. 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.

Automated summary

Peptide-assisted genome editing allows efficient editing of single and multiple genes in hematopoietic cells. This system enables simple and well-tolerated delivery of CRISPR-Cas system into primary cells with minimal toxicity. It achieves rapid and robust genome editing in primary cells, including T cells and hematopoietic progenitor cells, with editing efficiencies exceeding 98%. The Peptide-Assisted Genome Editing (PAGE) provides a versatile platform for next-generation genome engineering in primary cells.