Transduction of murine hematopoietic stem cells with tetracycline-regulated lentiviral vectors

Abstract

Tetracycline-regulated integrating vectors allow pharmacologically controlled genetic modifi cation of murine and human hematopoietic stem cells (HSCs). This approach combines the stable transgene insertion into a host genome with the opportunity for time- and dose-controlled reversible transgene expression in HSCs. Here, we describe the step-by-step protocol for transduction of murine stem-cell enriched populations of bone marrow cells, such as lineage negative cells (Lin −), with a lentiviral vector expressing the enhanced green fiuorescent protein (EGFP) under the control of the tetracycline-regulated promoter. This chapter explains how to establish in vitro and in vivo systems to study transgene dose-dependent mechanisms affecting cell fate decisions of genetically modifi ed hematopoietic cells.