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Anim Biotechnol. 2019 Feb 8:1-9. doi: 10.1080/10495398.2018.1559846. [Epub ahead of print]

Effect of supplemetation of Zebularine and Scriptaid on efficiency of in vitro developmental competence of ovine somatic cell nuclear transferred embryos.

Author information

1
a College of Animal Science and Technology , Hebei Agricultural University , Baoding , PR China.
2
b College of Life Science and Technology , Southwest Minzu University , Chengdu , China.
3
c State Key Laboratory for Sheep Genetic Improvement and Healthy Production, Institute of Animal Husbandry and Veterinary , Xinjiang Academy of Agricultural and Reclamation Sciences , Shihezi , China.
4
d College of Food Science and Technology , Hebei Agricultural University , Baoding , PR China.
5
e Animal Husbandry and Veterinary Research Institute of Tianjin , Tianjin , China.
6
f Research Center of Cattle and Sheep Embryo Engineering Technique of Hebei Province , Baoding , PR China.

Abstract

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.

KEYWORDS:

Scriptaid; Sheep; Zebularine; brilliant cresyl blue; somatic cell nuclear transfer

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