Xiong-XianRong, Lan-DaoLiang, Li Jian*, Zhong-JinCheng, Zi XiongDong, MaLi, Wang Yong.
CellReprogram,2013, 15(4): 293-300. (Impact factor 2.744, SCI)
Abstract: Abnormal epigenetic reprogramming of the donor nucleus after somatic cell nucleartransfer (SCNT) is thought to be the main cause of low cloning efficiency.Following SCNT, the donor nucleus of ten fails to express early embryonic genesand establish a normal embryonic pattern of chromatin modification. Therefore,in this study, we have attempted to improve epigenetic reprogramming of the donor nucleus and cloned embryos with Zebularine and Scriptaid. Yak fibroblasts were treated with 20μM Zebularine alone or 20μMZebularine plus 0.5μMScriptaid for 24 h, whereas yak cloned embryos were treated exclusively with 0.5μM Scriptaid for12 h. There was no effect on cellular viability and proliferation after drug treatment. The treatment of fibroblasts with Zebularine or Zebularine plusScriptaid increased histone acetylation of histone 3 lysine 9 (H3K9), but decreased the level of DNA methylation of Oct-4 and Sox-2 promoter regions.When donor cells were used after Zebularine plus Scriptaid treatment toreconstruct cloned embryos and then treated with Scriptaid, the developmental competence and cryosurvival of embryos were improved significantly. In addition, the relative expression of Oct-4 and significantly decreased when fibroblasts and cloned embryos were treated with Zebularine or Scriptaid. This work provides functional evidence that treatment with Zebularine and Scriptaid modifies the epigenetic status of yak fibroblasts, subsequently enhancing invitro developmental potential and the quality of yak cloned embryos.