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

Autor: Zhang, Zhen, Baxter, Amy E., Ren, Diqiu, Qin, Kunhua, Chen, Zeyu, Collins, Sierra M., Huang, Hua, Komar, Chad A., Bailer, Peter F., Parker, Jared B., Blobel, Gerd A., Kohli, Rahul M., Wherry, E. John, Berger, Shelley L., Shi, Junwei
Zdroj: Nature Biotechnology; Feb2024, Vol. 42 Issue 2, p305-315, 11p
Abstrakt: 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. Peptide-assisted genome editing enables efficient single and multiplex editing in hematopoietic cells. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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