T Bodies

Genetic modification of T cells with antigen-specific chimeric receptors (CARs) has great potential to redirect their effector functions to a wide variety of therapeutic targets, including weakly expressed or poorly presented tumor antigens. The protocol describes the subcloning of scFv fragments, costimulatory domains, and the T cell receptor (TCR) ζ chain, and their assembly as a chimeric T cell receptor gene. Furthermore, it provides an example for stable transfer of the recombinant receptor gene into activated human T cells.