Rhesus monkey placental transgene expression after lentiviral gene transfer into preimplantation embryos

Abstract

Transgenic mice have provided invaluable information about gene function and regulation. However, because of marked differences between rodents and primates, some areas of human biology such as early embryonic development, aging, and maternal-fetal interactions would be best studied in a nonhuman primate model. Here, we report that gene transfer into rhesus monkey (Macaca mulatta) preimplantation embryos gives rise to transgenic placentas that express a reporter transgene (eGFP). Blastocysts resulting from culture of in vitro fertilized ova were transduced with a selfinactivating lentiviral vector and transferred into recipient females. One twin and one singleton pregnancy were produced from a single stimulation cycle, and one live rhesus monkey was born from each pregnancy. Placentas from all conceptuses showed expression of the transgene as detected by reverse transcription-PCR, ribonuclease protection assay, direct epifluorescence, immunohistochemistry, and Western blot analysis. Integration in somatic tissues of the offspring was not detected. A maternal immune response to the xenogeneic placental antigen was shown by the presence of anti-GFP antibodies in peripheral blood of the recipient females by day 99 of gestation (term ‫؍‬ 165 days). These results demonstrate that transgene expression during gestation is compatible with successful pregnancy in nonhuman primates and provides an approach that could be broadly applicable to the development of novel models for primate biomedical research.

Efficient transgenesis in farm animals by lentiviral vectors

“…Further analyses revealed that the transgene was present only in extraembryonic tissues such as the placenta (Wolfgang et al, 2001). By contrast, we observed efficient transduction of the embryo proper and extraembryonic cells after infection of bovine embryos and oocytes.…”

“…The GFP transgene was not found in neonates developed from rhesus monkey embryos infected with lentiviral vectors (Wolfgang et al, 2001). Further analyses revealed that the transgene was present only in extraembryonic tissues such as the placenta (Wolfgang et al, 2001).…”

“…In addition, the lentiviral provirus was transmitted through the germ line. The different efficacies of lentiviral transgenesis in monkey versus livestock might be due to basic biological differences as well as technical aspects such as the time-point of virus injection (single-cell embryos versus blastocysts; Wolfgang et al, 2001) and virus titre. An important feature of lentiviral transgenesis in livestock is the linear correlation of transgene expression and number of integrants.…”

“…However, attempts to generate transgenic monkeys by lentiviral infection of pre-implantation embryos were not successful and resulted in gene transfer only into extraembryonic tissues (Wolfgang et al, 2001), raising the question whether lentiviral vectors can be used for transgenesis in higher mammals.…”

Toll-like receptor 7 and MyD88 knockdown by lentivirus-mediated RNA interference to porcine dendritic cell subsets

“…[31][32][33][34] These viruses have also been employed successfully to deliver exogenous genes to human cDC. [35][36][37][38] A major advantage of this vector system is its capacity to mediate stable foreign gene expression in non-dividing cells -a prerequisite for studies on DC activity.…”