RepSox improves viability and regulates gene expression in rhesus monkey–pig interspecies cloned embryos
To investigate the effect of the small molecule, RepSox, on the expression of developmentally important genes and the pre-implantation development of rhesus monkey–pig interspecies somatic cell nuclear transfer (iSCNT) embryos.
Rhesus monkey cells expressing the monomeric red fluorescent protein 1 which have a normal (42) chromosome complement, were used as donor cells to generate iSCNT embryos. RepSox increased the expression levels of the pluripotency-related genes, Oct4 and Nanog (p < 0.05), but not of Sox2 compared with untreated embryos at the 2–4-cell stage. Expression of the anti-apoptotic gene, Bcl2, and the pro-apoptotic gene Bax was also affected at the 2–4-cell stage. RepSox treatment also increased the immunostaining intensity of Oct4 at the blastocyst stage (p < 0.05). Although the blastocyst developmental rate was higher in the group treated with 25 µM RepSox for 24 h than in the untreated control group (2.4 vs. 1.2%, p > 0.05), this was not significant.
RepSox can improve the developmental potential of rhesus monkey–pig iSCNT embryos by regulating the expression of pluripotency-related genes.
KeywordsEmbryos (monkey–pig) Interspecies (monkey–pig) somatic cell nuclear transfer mRFP1 Pluripotency RepSox Rhesus monkey
We thank Xiaohui Wu at Fudan University for providing the pCX-mRFP1 vector. This work was supported by the Science and Technology Development Projects of Jilin Province of China (Grant No. 20160520057JH) and the 13th Five-Year Plan Science and Technology Research Project of Education Department of Jilin Province of China (Grant No. -257).
Supplementary Table 1—List of primers.