Journal article
CRISPitope: A generic platform to model target antigens for adoptive T cell transfer therapy in mouse tumor models
M Effern, N Glodde, E Bawden, J Liebing, D Hinze, T Tüting, T Gebhardt, M Hölzel
STAR Protocols | Published : 2022
Abstract
This protocol details the procedure for CRISPR-assisted insertion of epitopes (CRISPitope), a flexible approach for generating tumor cells expressing model CD8+ T cell epitopes fused to endogenously encoded gene products of choice. CRISPitope-engineered tumor cells can be recognized by T cell receptor-transgenic (TCRtg) CD8+ T cells that are widely used in immunology research. Using mice inoculated with CRISPitope-engineered tumor cells, researchers can investigate how the choice of the target antigen for T cell immunotherapies influences treatment efficacy and resistance mechanisms. For complete details on the use and execution of this protocol, please refer to Effern et al. (2020).
Grants
Awarded by Deutsche Forschungsgemeinschaft
Funding Acknowledgements
We thank P. Wurst and An. Dolf for help with flow cytometry. We thank the UKB core facility flow cytometry. This work was supported by the Deutsche Krebshilfe (German Cancer Aid) through a junior research group grant within the Mildred Scheel School of Oncology (MSSO) Cologne-Bonn (70113307) to N.G., the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) within GRK 2168 to M.H., scholarships by the University of Melbourne to M.E. and E.B., a scholarship to M.E. within GRK 2168, and the DFG under Germany's Excellence Strategy-EXC2151-390873048. T.G. and M.H. are supervisors in the GRK 2168 international graduate school, Bonn-Melbourne. M.H. isa member of the DFG Excellence Cluster ImmunoSensation2 (EXC 2151). We thank D. Cor-vino, S. Ng, and M. Yong from the Institute of Experimental Oncology for critical reading. The graph-ical abstract, parts of Figures 2 and 5 were generated using biorender.com. Figure 3 was generated using SnapGene software (from insightful Science; available at snapgene.com). Figures 1, 4A, 4C, and 6 are reprinted from (Effern et al., 2020), with permission from Elsevier.