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  • Open Access

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

  • 1,
  • 1 and
  • 2
Journal for ImmunoTherapy of Cancer20153 (Suppl 2) :P53

  • Published:


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


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


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


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.


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.



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).

Authors’ Affiliations

The Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Nanjing, China
The Comprehensive Cancer Center of Drum-Tower Hospital, Medical School of Nanjing University & Clinical Cancer Institute of Nanjing University, Nanjing, China


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© Su et al. 2015

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.