Genetic in vivo engineering of human T lymphocytes in mouse models

Abstract Receptor-targeting of vector particles is a key technology for cell-type specific in vivo gene delivery. Lentiviral vectors (LVs) targeted to human T cell markers enable genetic modification of T cells directly in humanized mouse models. Illustrating the example of modifying T cells with chimeric antigen receptors (CARs), we provide detailed protocols for the generation and quality check of potent CD4- and CD8-targeted LVs, the humanization of immunodeficient mice as well as the administration of the vector stocks followed by monitoring for in vivo modified T cells. By closely following the protocol, sufficient vector stock for the genetic manipulation of 10-15 humanized mice is obtained. A few weeks after administration, about 10% of T cells can be expected to convert to CAR T cells. The protocol can be easily adapted to LVs targeted to other types of receptors and/or delivering other genes of interest.