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TCR and CAR engineering of primary human T cells

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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