Folic Acid Deprivation and siRNA-Inhibition of Su-v39h1/2 in Fibroblast Donor Cells Improves Reprogram-ming of Bovine SCNT Embryos

Publish Year: 1398
نوع سند: مقاله کنفرانسی
زبان: English
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RROYAN20_153

تاریخ نمایه سازی: 29 مهر 1398

Abstract:

Efficiency of SCNT has remained low due to a strong resistance of somatic donor cells to epigenetic reprogramming. Many epigenetic drugs, targeting epigenetic status of donor cells or reconstructed oocytes to improve development of SCNT em-bryos. In our recent studies, we examined the effect of siRNA inhibition and chemically inhibition of H3K9me3 and also induced DNA hypo-methylation following in vitro folate de-ficiency in fibroblast donor cells on in vitro development of bovine SCNT embryos. Chaetocin was supplemented during the culture of donor cells for 3 days. In addition siRNA knock-down of SUV39H1/H2 was done in donor cells. Both chaetocin and siSUV39H1/H2 significantly reduced the relative intensity level of H3K9me3 in fibroblast cells. siSUV39H1/H2 transfec-tion but not chaetocin treatment improved in vitro development of SCNT embryos. In addition, siSUV39H1/H2 altered the ex-pression profile of the selected genes in the derived blastocysts similar to those derived from IVF. DNA methylation in cells cultured in folate deficient (folate -) medium in presence of 0.5% serum was decreased. Bisulfite sequencing analysis in-dicated a decrease in DNA methylation of POU5F1 promoter and gene expression analysis revealed an increase in expres-sion of POU5F1 gene. Blastocyst rate in folate - group was significantly higher than folate + group. The DNA methylation level in POU5F1 promoter and ICR of H19 and IGF2 of SCNT derived embryos in folate - group was similar to IVF derived blastocysts. In conclusion, our results may provide two new and novel approaches for improving mammalian SCNT efficiency for agricultural and biomedical purposes.

Authors

F Jafarpour

Department of Reproductive Biotechnology, Reproductive Bio-medicine Research Center, Royan Institute for Biotechnology,ACECR, Isfahan, Iran