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

Maike Effern; Nicole Glodde; Emma Bawden; Jana Liebing; Daniel Hinze; Thomas Tüting; Thomas Gebhardt; Michael Hölzel

doi:10.1016/j.xpro.2021.101038

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

STAR Protoc. 2022 Jan 7;3(1):101038. doi: 10.1016/j.xpro.2021.101038. eCollection 2022 Mar 18.

Authors

Maike Effern^{1

2}, Nicole Glodde¹, Emma Bawden^{1

2}, Jana Liebing¹, Daniel Hinze¹, Thomas Tüting³, Thomas Gebhardt², Michael Hölzel¹

Affiliations

¹ Institute of Experimental Oncology (IEO), Medical Faculty, University Hospital Bonn, University of Bonn, 53105 Bonn, Germany.
² Department of Microbiology & Immunology, The University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, VIC, Australia.
³ Laboratory of Experimental Dermatology, Department of Dermatology, University of Magdeburg, Magdeburg, Germany.

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).

Keywords: Biotechnology and bioengineering; CRISPR; Cancer; Cell Biology; Flow Cytometry/Mass Cytometry; Immunology; Model Organisms; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CD8-Positive T-Lymphocytes*
Disease Models, Animal
Epitopes, T-Lymphocyte
Immunotherapy, Adoptive / methods
Mice
Neoplasms* / therapy
Receptors, Antigen, T-Cell / genetics

Substances

Epitopes, T-Lymphocyte
Receptors, Antigen, T-Cell