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CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells for adoptive therapy

Background

Strategies that enhance the function of T cells are critical for immunotherapy.

Methods

Here we described for the first time a non-viral mediated approach to reprogram primary human T cells by disruption of PD-1.

Results

We showed that the gene knockout of PD-1 by electroporation of plasmids encoding sgRNA and Cas9 was technically feasible. The disruption of PD-1 resulted in significant reduction of PD-1 expression but didn't affect the viability of primary human T cells. Cellular immune response of the gene modified T cells was characterized by up-regulated IFN-γ production and enhanced cytotoxicity.

Conclusions

These results suggest that we have established an approach for efficient checkpoint inhibitor disruption, providing a new strategy for targeting checkpoint inhibitors to improve the efficacy of T cell based adoptive therapies.

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Acknowledgements

The authors thank the entire Huang Lab and Liu Lab for their support and advice. This work was funded by grants from the National Natural Science Foundation of China (Grant No. 81172281, 81000980, 81220108023, 81172094).

Author information

Correspondence to Zhengyun Zou.

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Keywords

  • Public Health
  • Immune Response
  • Cellular Immune Response
  • Gene Knockout
  • Mediate Approach