Journal
NATURE PROTOCOLS
Volume 15, Issue 4, Pages 1507-1524Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41596-020-0294-8
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Funding
- National Institutes of Health [DP5OD012133]
- National Institutes of Health (UC CAI) [U54HL119893]
- National Institutes of Health (UCLA CTSI) [UL1TR001881]
- NIH F30 Fellowship [F30CA183528]
- NIH Biotechnology Training in Biomedical Sciences and Engineering Program [T32 GM067555]
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The expression of synthetic receptors in primary T cells enables the programming of user-defined responses when designing T-cell therapies. Chimeric antigen receptors (CARs) are synthetic receptors that have demonstrated efficacy in cancer therapy by targeting immobilized antigens on the surface of malignant cells. Recently, we showed they can also rewire T-cell responses to soluble ligands. In contrast to other synthetic receptors, CARs are not only readily engineered by rational design, but also clinically translatable, with robust function in primary human T cells. This protocol discusses design principles for CARs responsive to soluble ligands and delineates steps for producing T cells expressing synthetic receptors. Functional assays for quantifying the ability of CAR T cells to sense and respond to soluble ligands are also presented. This protocol provides a framework for proficient immune-cell researchers to test novel T-cell therapies targeting soluble ligands in <2 weeks. This protocol discusses design principles for CARs responsive to soluble ligands and delineates steps for producing T cells expressing synthetic receptors. Functional assays for quantifying the ability of CAR T cells to sense and respond to soluble ligands are also presented.
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