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| Item Type: | Article |
|---|---|
| Title: | TCR and CAR engineering of primary human T cells |
| Creators Name: | Edes, I., Clauss, J., Stahn, R., Sada Japp, A. and Lorenz, F.K.M. |
| Abstract: | The efficient expression of T-cell receptors (TCRs) or chimeric antigen receptors (CARs) in primary human T cells is crucial for preclinical testing of receptor properties for adoptive T-cell therapies. Multiple streams of technological platforms have been developed in the recent decades to genetically modify primary T cells including nonviral platforms such as transposon-based systems (PiggyBac, Sleeping Beauty), TALENs, or CRISPR-Cas9). The production of CAR- or TCR-encoding retroviral vectors, however, is still the most commonly used technique both in preclinical as well as in clinical settings.In this chapter we describe a comprehensive 12-day protocol for (a) generating high-titered gamma-retroviral vector particles containing the transgene of interest (e.g., TCR , CAR ), (b) the isolation, activation and rapid expansion of primary T cells and (c) the stable genetic engineering of these T cells with the transgene for subsequent characterization. |
| Keywords: | Primary Human T Cells, T-Cell Receptor, Chimeric Antigen Receptor, Gamma-Retrovirus, Retroviral Vector, Transfection, Transduction, Genetic Engineering |
| Source: | Methods in Molecular Biology |
| Series Name: | Methods in Molecular Biology |
| Title of Book: | Gene therapy of cancer : methods and protocols |
| ISSN: | 1064-3745 |
| ISBN: | 9781071624401 |
| Publisher: | Springer / Humana Press |
| Volume: | 2521 |
| Number: | 3 |
| Page Range: | 85-93 |
| Number of Pages: | 445 |
| Date: | 23 June 2022 |
| Official Publication: | https://doi.org/10.1007/978-1-0716-2441-8_5 |
| PubMed: | View item in PubMed |
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