Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair
Summary: 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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721007371 |
| _version_ | 1818646579151634432 |
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| author | Carolin Moosmann Thomas R. Müller Dirk H. Busch Kilian Schober |
| author_facet | Carolin Moosmann Thomas R. Müller Dirk H. Busch Kilian Schober |
| author_sort | Carolin Moosmann |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-17T00:48:53Z |
| format | Article |
| id | doaj.art-578d4813e24a42b3b9fc77072066d5e6 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-17T00:48:53Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-578d4813e24a42b3b9fc77072066d5e62022-12-21T22:09:48ZengElsevierSTAR Protocols2666-16672022-03-0131101031Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repairCarolin Moosmann0Thomas R. Müller1Dirk H. Busch2Kilian Schober3Mikrobiologisches Institut–Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany; Corresponding authorInstitute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, Munich, GermanyInstitute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, Munich, GermanyMikrobiologisches Institut–Klinische Mikrobiologie, Immunologie und Hygiene, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166721007371Health SciencesImmunologyCRISPRBiotechnology and bioengineering |
| spellingShingle | Carolin Moosmann Thomas R. Müller Dirk H. Busch Kilian Schober Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair STAR Protocols Health Sciences Immunology CRISPR Biotechnology and bioengineering |
| title | Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair |
| title_full | Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair |
| title_fullStr | Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair |
| title_full_unstemmed | Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair |
| title_short | Orthotopic T-cell receptor replacement in primary human T cells using CRISPR-Cas9-mediated homology-directed repair |
| title_sort | orthotopic t cell receptor replacement in primary human t cells using crispr cas9 mediated homology directed repair |
| topic | Health Sciences Immunology CRISPR Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721007371 |
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