[Development and evaluation of a novel placenta-specific gene manipulation method using lentiviral vectors]

The placenta plays numerous important roles to support fetal development such as gas exchange, nutrient supply, and hormone production. Placental defects underlie many aspects of pregnancy losses and complications; thus understanding and regulating gene function during placentation is of high clinical relevance. However, the lack of a facile and efficient method for placenta-specific gene manipulation has hampered study of the placenta. We have previously shown that transduction of fertilized mouse eggs with lentiviral (LV) vectors efficiently generates transgenic animals; however, transgene expression occurred in both the fetus and the placenta. In the present study, we transduced zona-free blastocysts with LV vectors expecting placenta-specific gene expression, since most placental cells differentiate from trophoblast cells that form the outermost layer of the blastocyst. Transgene expression was observed in trophoblast cells from preimplantation stages and in the placenta throughout gestation. All the analyzed placentas carried the transgene, while none of the fetuses became transgenic. By applying this method, embryonic lethality caused by placental defects in several knockout animal models was substantially rescued. This technology provides a powerful system for gene manipulation exclusively in placental organogenesis with implications for the treatment of placental dysfunction.