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. and Clauss, J. and Stahn, R. and 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 |
---|
Related to: | |
---|
Repository Staff Only: item control page
|