Journal article

Efficient CRISPR/Cas9 Gene Editing in Uncultured Naive Mouse T Cells for In Vivo Studies

Simone Nussing, Imran G House, Conor J Kearney, Amanda XY Chen, Stephin J Vervoort, Paul A Beavis, Jane Oliaro, Ricky W Johnstone, Joseph A Trapani, Ian A Parish

The Journal of Immunology | AMER ASSOC IMMUNOLOGISTS | Published : 2020

Abstract

CRISPR/Cas9 technologies have revolutionized our understanding of gene function in complex biological settings, including T cell immunology. Current CRISPR-mediated gene editing strategies in T cells require in vitro stimulation or culture that can both preclude the study of unmanipulated naive T cells and alter subsequent differentiation. In this study, we demonstrate highly efficient gene editing within uncultured primary naive murine CD8+ T cells by electroporation of recombinant Cas9/sgRNA ribonucleoprotein immediately prior to in vivo adoptive transfer. Using this approach, we generated single and double gene knockout cells within multiple mouse infection models. Strikingly, gene deleti..

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Grants

Awarded by Human Frontier Science Program


Funding Acknowledgements

This work was supported by Human Frontier Science Program Young Investigators Grant RGY0065/2018 and by program, fellowship, and project grant support from the National Health and Medical Research Council of Australia.