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

Carolin Moosmann, Thomas R. Müller, Dirk H. Busch, Kilian Schober

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

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).

Original languageEnglish
Article number101031
JournalSTAR Protocols
Volume3
Issue number1
DOIs
StatePublished - 18 Mar 2022
Externally publishedYes

Keywords

  • Biotechnology and bioengineering
  • CRISPR
  • Health Sciences
  • Immunology

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