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| 001 | 163037 | ||
| 005 | 20240229123146.0 | ||
| 024 | 7 | _ | |a 10.1371/journal.pone.0238875 |2 doi |
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| 037 | _ | _ | |a DKFZ-2020-01828 |
| 041 | _ | _ | |a eng |
| 082 | _ | _ | |a 610 |
| 100 | 1 | _ | |a Kropp, Korbinian N |0 0000-0001-8104-7166 |b 0 |
| 245 | _ | _ | |a A bicistronic vector backbone for rapid seamless cloning and chimerization of αβT-cell receptor sequences. |
| 260 | _ | _ | |a San Francisco, California, US |c 2020 |b PLOS |
| 336 | 7 | _ | |a article |2 DRIVER |
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| 520 | _ | _ | |a To facilitate preclinical testing of T-cell receptors (TCRs) derived from tumor-reactive T-cell clones it is necessary to develop convenient and rapid cloning strategies for the generation of TCR expression constructs. Herein, we describe a pDONR™221 vector backbone allowing to generate Gateway™ compatible entry clones encoding optimized bicistronic αβTCR constructs. It harbors P2A-linked TCR constant regions and head-to-head-oriented recognition sites of the Type IIS restriction enzymes BsmBI and BsaI for seamless cloning of the TCRα and TCRβ V(D)J regions, respectively. Additional well-established TCR optimizations were incorporated to enhance TCR functionality. This included replacing of the human αβTCR constant regions with their codon-optimized murine counterparts for chimerization, addition of a second interchain disulfide bond and arrangement of the TCR chains in the order β-P2A-α. We exemplified the utility of our vector backbone by cloning and functional testing of three melanoma-reactive TCRs in primary human T cells. |
| 536 | _ | _ | |a 314 - Tumor immunology (POF3-314) |0 G:(DE-HGF)POF3-314 |c POF3-314 |f POF III |x 0 |
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| 700 | 1 | _ | |a Schäufele, Tim J |b 1 |
| 700 | 1 | _ | |a Fatho, Martina |b 2 |
| 700 | 1 | _ | |a Volkmar, Michael |0 P:(DE-He78)792fd96345efded9a78c902105479a68 |b 3 |u dkfz |
| 700 | 1 | _ | |a Conradi, Roland |b 4 |
| 700 | 1 | _ | |a Theobald, Matthias |b 5 |
| 700 | 1 | _ | |a Wölfel, Thomas |b 6 |
| 700 | 1 | _ | |a Wölfel, Catherine |b 7 |
| 773 | _ | _ | |a 10.1371/journal.pone.0238875 |g Vol. 15, no. 9, p. e0238875 - |0 PERI:(DE-600)2267670-3 |n 9 |p e0238875 - |t PLOS ONE |v 15 |y 2020 |x 1932-6203 |
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| 914 | 1 | _ | |y 2020 |
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