Non-viral targeted integration of large DNA in primary human T cells independent of double-stranded DNA breaks

Apr 30, 2026Nature biomedical engineering

Precise insertion of large DNA into human T cells without cutting their DNA

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Abstract

PRIME-In achieves up to 50% integration efficiency for a 3-kb CAR construct in human primary T cells.

  • PRIME-In utilizes a novel genome-editing platform that integrates large DNA cargoes without causing genomic double-strand breaks.
  • This method enhances editing efficiency and specificity compared to traditional DSB-dependent techniques.
  • PRIME-In eliminates detectable on-target and off-target chromosomal abnormalities during the editing process.
  • Refinements in reagent composition and delivery protocols have led to minimal toxicity in the engineering of primary human T cells.
  • The platform may offer significant advancements in the non-viral production of genome-edited T cells for immunotherapy applications.

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