Exploiting hypoxia in solid malignancies to restrict expression of chimeric antigen receptors (CARs) on engineered T cells to the tumor microenvironment overcomes the risk of on-target off-tumor toxicity and minimizes tonic signaling, which promotes CAR T cell exhaustion. This protocol summarizes the synthetic biology underlying the development of a stringent oxygen-sensitive CAR for in vitro and in vivo preclinical characterization. For complete details on the use and execution of this protocol, please refer to Kosti et al. (2021).
Keywords: Biotechnology and bioengineering; Cancer; Cell culture; Cell isolation; Cell separation/fractionation; Cell-based assays; Flow cytometry/mass cytometry; Immunology; Molecular biology.
© 2021 The Author(s).