Lentiviral Vector-Mediated Gene Transfer in Embryonic Stem Cells

The major limitations in gene transduction to embryonic stem (ES) cells are (1) low efficiency of gene delivery and (2) suppression of gene expression after integration into the host genome. A human immunodeficiency virus type 1 (HIV-1)-based lentiviral vector has been demonstrated to be an excellent tool for stable and efficient gene expression in ES cells. Here, we introduce a protocol for lentiviral vector-mediated transgene expression in murine ES cells. Using lentiviral vectors expressing LacZ, green fluorescent protein, and Cre recombinase, we demonstrate the efficiency and utility of the vectors in ES cell study.