Effect of supplemetation of Zebularine and Scriptaid on efficiency of in vitro developmental competence of ovine somatic cell nuclear transferred embryos.
Somatic cell nuclear transfer (SCNT) technology has been applied in the construction of disease model, production of transgenic animals, therapeutic cloning, and other fields. However, the cloning efficiency remains limited. In our study, to improve SCNT efficiency, brilliant cresyl blue (BCB) staining were chosen to select recipient oocytes. In addition, DNA methyltransferase inhibitor Zebularine (5 nmol/L) and histone deacetylase inhibitor Scriptaid (0.2 μmol/L) were jointly used to treat sheep donor cumulus cells and reconstructed embryo. Moreover, the expression levels of embryonic development-related genes (OCT4, SOX2, H19, IGF2 and Dnmt1) of reconstructed embryo were also detected. Using BCB + oocytes as recipient cell, donor cumulus cells and reconstructed embryos were treated with 5 nmol/L Zebularine and 0.2 μmol/L Scriptaid, the blastocyst rate in Zeb + SCR-SCNT group (28.25%) was significantly higher than SCNT (21.16%) (p < 0.05). Furthermore, results showed that expression levels of OCT4, SOX2, H19, IGF2 and Dnmt1 genes in Zeb + SCR-SCNT embryos were more similar to IVF embryos. Our study proved that 5 nmol/L Zebularine and 0.2 μmol/L Scriptaid treating with sheep donor cumulus cells and reconstructed embryos improved SCNT blastocyst rate and relieve the abnormal expression of embryonic developmental related genes.
['Fang|Xiaohuan|X|', 'Xia|Wei|W|', 'Cao|Hui|H|', 'Guo|Yanhua|Y|', 'Wang|Han|H|', 'Zhang|Xiaosheng|X|', 'Wan|Pengcheng|P|', 'Liu|Chuang|C|', 'Wei|Qiaoli|Q|', 'Sun|Shuchun|S|', 'Tian|Shujun|S|', 'Li|Junjie|J|', 'Wang|Zhigang|Z|']
D000818:Animals D019976:Cloning, Organism D003562:Cytidine D004622:Embryo, Mammalian D047108:Embryonic Development D018507:Gene Expression Regulation, Developmental D056572:Histone Deacetylase Inhibitors D006898:Hydroxylamines D053652:Nuclear Transfer Techniques D011804:Quinolines D012756:Sheep
Zebularine Scriptaid somatic cell nuclear transfer brilliant cresyl blue Sheep