Protocol for ecotropic pseudotyped lentiviral transduction of “murinized” human cells while decreasing risk of infection for laboratory personnel

Lentiviral systems of transduction are valuable tools to stably express transgenes of interest in human cell lines. However, their use risks infection of the lab workers. Here, we present a protocol for lentiviral vector system pseudotyping with the murine ecotropic envelope. We detail steps for producing ecotropic pseudotyped lentiviral particles and transiently expressing the Slc7a1 receptor for murine leukemia virus in target human cells. We then describe procedures for puromycin selection and the characterization of stable cell lines for transgene expression. For complete details on the use and execution of this protocol, please refer to Iyer-Bierhoff et al.¹.