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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Vydáno v:	STAR protocols Ročník 3; číslo 1; s. 101038
Hlavní autoři:	Effern, Maike, Glodde, Nicole, Bawden, Emma, Liebing, Jana, Hinze, Daniel, Tüting, Thomas, Gebhardt, Thomas, Hölzel, Michael
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	United States Elsevier Inc 18.03.2022 Elsevier
Témata:	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
On-line přístup:	Získat plný text
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Popis
Shrnutí:	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.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Technical contact Lead contact
ISSN:	2666-1667 2666-1667
DOI:	10.1016/j.xpro.2021.101038