CRISPitope: A generic platform to model target antigens for adoptive T cell transfer therapy in mouse tumor models

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 recept...

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Veröffentlicht in:STAR protocols Jg. 3; H. 1; S. 101038
Hauptverfasser: Effern, Maike, Glodde, Nicole, Bawden, Emma, Liebing, Jana, Hinze, Daniel, Tüting, Thomas, Gebhardt, Thomas, Hölzel, Michael
Format: Journal Article
Sprache:Englisch
Veröffentlicht: United States Elsevier Inc 18.03.2022
Elsevier
Schlagworte:
Animals
Biotechnology and bioengineering
Cancer
CD8-Positive T-Lymphocytes
Cell Biology
CRISPR
Disease Models, Animal
Epitopes, T-Lymphocyte
Flow Cytometry/Mass Cytometry
Immunology
Immunotherapy, Adoptive - methods
Mice
Model Organisms
Molecular Biology
Neoplasms - therapy
Protocol
Receptors, Antigen, T-Cell - genetics
CRISPR
Immunology
Molecular Biology
Biotechnology and bioengineering
Flow Cytometry/Mass Cytometry
Model Organisms
Cell Biology
Cancer
ISSN:2666-1667, 2666-1667
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Zusammenfassung: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). [Display omitted] •Genome editing of tumor cells for experimental standardized T cell recognition•Tagging endogenous gene products with model CD8+ T cell epitopes in tumor cells•Assessing T cell responses directed against differentially regulated gene products•Approach allows studying the impact of the target antigen choice on T cell therapy 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.
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ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2021.101038