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 (5nmol/L) and histone deacetylase inhibitor Scriptaid (0.2 mu mol/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 BCBthornoocytes as recipient cell, donor cumulus cells and reconstructed embryos were treated with 5nmol/L Zebularine and 0.2 mu mol/L Scriptaid, the blastocyst rate in ZebthornSCR-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 ZebthornSCR-SCNT embryos were more similar to IVF embryos. Our study proved that 5nmol/L Zebularine and 0.2 mu mol/ 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.