Journal
JOURNAL OF EXPERIMENTAL MEDICINE
Volume 219, Issue 5, Pages -Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1084/jem.20211530
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The researchers report a convenient and scalable nonviral method for precise gene editing and transgene integration in primary human T cells. This method is valuable for advancing our understanding of T cell biology and the development of next-generation cellular therapies.
Genome engineering of T lymphocytes, the main effectors of antitumor adaptive immune responses, has the potential to uncover unique insights into their functions and enable the development of next-generation adoptive T cell therapies. Viral gene delivery into T cells, which is currently used to generate CAR T cells, has limitations in regard to targeting precision, cargo flexibility, and reagent production. Nonviral methods for effective CRISPR/Cas9-mediated gene knock-out in primary human T cells have been developed, but complementary techniques for nonviral gene knock-in can be cumbersome and inefficient. Here, we report a convenient and scalable nonviral method that allows precise gene edits and transgene integration in primary human T cells, using plasmid donor DNA template and Cas9-RNP. This method is highly efficient for single and multiplex gene manipulation, without compromising T cell function, and is thus valuable for use in basic and translational research. Effective and precise gene editing of T lymphocytes is critical for advancing our understanding of T cell biology and the development of next-generation cellular therapies. The authors report a simple and efficient method for nonviral CRISPR/Cas9-based editing of T lymphocytes using plasmid donor DNA.
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