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Generation of hypoxia-sensing chimeric antigen receptor T?cells

STAR Protoc. 2021-08; 
Paris Kosti, Karen I Larios-Martinez, John Maher, James N Arnold
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Gene Synthesis … Note: Synthetic DNA fragments can be ordered from Integrated DNA Technologies (IDT), GenScript, or others?… HRE synthetic DNA fragment (1,480 bp) together using T4 DNA Ligase, at a molar ratio of 3:1 insert to vector, according to manufacturers' protocol: https://international?… Get A Quote

摘要

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 and preclinical characterization. For complete details on the use and execution of this protocol, please refer to Kosti et?al. (2021).

關鍵詞

Biotechnology and bioengineering, Cancer, Cell culture, Cell isolation, Cell separation/fractionation, Cell-based assays, Flow cytometry/mass cytometry, Immunology, Molecular biology
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