Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair.

Moosmann C, Müller TR, Busch DH, Schober K (2022)

Publication Status: Published

Publication Type: Journal article

Publication year: 2022

Journal

STAR Protocols Cell Press

Book Volume: 3

Pages Range: 101031

Journal Issue: 1

DOI: 10.1016/j.xpro.2021.101031

Abstract

Adoptive T cell therapy using T-cell receptor (TCR)-engineered T cells allows to redirect T cell specificity and to target any antigen of interest. Here, we apply advanced genetic engineering using clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) for simultaneous editing of TCR α- and β-chains in primary human T cells. Together with non-virally delivered template DNA, this CRISPR-Cas9-system allows for elimination of the endogenous TCR and orthotopic placement of TCR α- and β-chains. For complete details on the use and execution of this protocol, please refer to Schober et al. (2019) and Müller et al. (2021).

Authors with CRIS profile

Carolin Moosmann Institute of Microbiology – Clinical Microbiology, Immunology and Hygiene Kilian Schober Institute of Microbiology – Clinical Microbiology, Immunology and Hygiene

Involved external institutions

Technische Universität München (TUM)

Germany (DE)

How to cite

APA:

Moosmann, C., Müller, T.R., Busch, D.H., & Schober, K. (2022). Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair. STAR Protocols, 3(1), 101031. https://doi.org/10.1016/j.xpro.2021.101031

MLA:

Moosmann, Carolin, et al. "Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair." STAR Protocols 3.1 (2022): 101031.

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