Methods and Reagents for The Generation of Chimeric Antigen Receptors Using Split in Teins

The use of chimeric antigen receptors (CARs) against fixed tumor-associated antigens (TAAs) is limited by the use of one CAR for each single therapy. So new strategies are required to develop flexible and modular CAR T systems, allowing for multiple antigens targeting.Chimeric antigen receptros (CARs) are used in many clinical applications, including cancer treatment. A chimeric antigen receptor is a recombinant receptor composed of an extracellular antigen binding domain and an intracellular T-cell signalling domain. The use of chimeric antigen receptor (CAR)-modified T cells is an innovative immunotherapeutic approach. CAR cell therapy relies on re-engineering T-cells to express a receptor that allowe the cells to recognize targeted cells. Typically, CAR treatment includes collecting T cells from a patient and introducing a chimeric antigen into the collected cells ex vivo, expanding the transfected cells, and then infusing them into a patient.The present invention relates to methods and reagents for the generation of Chimeric Antigen Receptors using split inteins as well as to the uses of the Chimeric Antigen Receptors, particularly in disease requiring a cell-mediated immune response against an antigen of interest. The present invention also relates to caged inteins and methods for the generation of recombinant proteins, in particular, chimeric antigen receptors using the caged inteins.