Abstract: A nucleic acid construct for targeting and integrating a CD3 zeta-deficient chimeric antigen receptor (CAR) fragment into an endogenous CD3 zeta/CD247 gene of a host genome. Also disclosed is a genetically modified human cell expressing an exogenous nucleic acid sequence encoding a CD3 zeta deficient CAR fragment, integrated in-frame into the endogenous CD3 zeta/CD247 gene for gene fusion, to form a functional CAR including an exogenous CAR fragment fused with an endogenous CD3 zeta domain.

Abstract: The present invention provides to the field of cell therapy, in particular, to adoptive T cell or NK cell therapy. It provides a human immunophilin knockout cell that is a T cell or NK cell, wherein the immunophilin is FKBP12 or cyclophilin A, or a cell population of such cells. The cell is resistant to the action of the immunosuppressant Tacrolimus and/or cyclosporine A. It is obtainable by CRISPR/Cas, e.g., CRISPR/Cas9-mediated knockout of the immunophilin or a derivate technology. The cell can be, e.g., a virus-specific cell, e.g., specific for an antigen from any of CMV, EBV, BKV, HPV, ADV, influenza virus, or SARS-CoV-2. It may also be a regulatory T cell.