RepSox improves viability and regulates gene expression in rhesus monkey–pig interspecies cloned embryos
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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.