Characteristics of induced pluripotent stem (iPS) cells are consistent with those of embryonic stem (ES) cells. However, exogenous genes integrated by using retrovirus delivery systems cannot be completely removed from the cells. In a recent report, activation-induced cytosine deaminase (AID) and thymine DNA glycosylase (TDG) can induce pluripotency state in mouse differentiated cells through the process of DNA demethylation. Thus, we hypothesized that the two reprogramming factors may convert efficiently bovine differentiated cells into pluripotency state. So, genes of AID and TDG were integrated into pCMV6-AC-IRES-GFP-Puro expression vector, which was transfected into bovine differentiated cells. As results, the colonies derived from AID+TDG-induced bovine cells were formed on day 7 after culture. The number of AP positively colonies in AID+TDG-induced bovine cells was significantly higher than in AID-induced bovine cells (p<0.05). Additionally, expression of pluripotent genes (OCT-3/4, NANOG, SOX2) was slightly increased in AID+TDG-induced bovine cells, as compared to AID-induced bovine cells. Protein expressions of OCT-3/4, NANOG and SOX2 in AID+TDG-induced bovine cells were slightly increased rather than AID-induced bovine cells. Finally, DNA demethylation in the promoter regions of pluripotent markers in AID+TDG-induced bovine cells was increased than that of AID-induced bovine cells. In conclusion, pluripotent stem cells could be efficiently produced from bovine differentiated cells by using non-integrating delivery system with the reprogramming factors (AID and TDG).

The companion cells of the Arabidopsis thaliana egg and sperm, the central and vegetative cells, undergo active DNA demethylation prior to fertilization. However, its biological significance, extent of conservation, and targeting preferences are not yet clear. We recently showed that localized demethylation of interspersed, small transposable elements is a common feature of A. thaliana companion cells. The DEMETER DNA glycosylase encodes active DNA demethylase activity and is required for seed production. DME-mediated DNA demethylation in the central cell is required to establish imprinted gene expression in the endosperm, and is considered a master regulator for plant gene imprinting. However, the similarity among DME targets in the central and vegetative cells, despite their different functions and developmental fates, suggests that establishment of genomic imprinting may not be the basal function of DME. Lack of DEMETER in vegetative cells causes reduced methylation of transposons in sperm. Our observation suggests that the primary function of companion cell demethylation is to reinforce transposon silencing in plant gametes.