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 I (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.
Oka
Masahiro
M
Department of Pathology, University of Florida College of Medicine, Gainesville, FL, USA.
Chang
Lung-Ji
LJ
Costantini
Frank
F
Terada
Naohiro
N
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D004622:Embryo, Mammalian / Q000166:cytology*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D019426:Integrases / Q000235:genetics; D007763:Lac Operon; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D011995:Recombination, Genetic; D014161:Transduction, Genetic
D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases