Abundance of TRIM28, SETDB1, and TP53 mRNA is dynamically regulated during porcine early embryogenesis and is abnormal in preimplantation embryos produced by in vitro fertilization in comparison to in vivo derived and nuclear transfer derived embryos
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[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] In vitro embryo production is important for research in animal reproduction, embryo transfer, transgenics, and cloning. However, in vitro-derived embryos generally have delayed development and are inferior to in vivo-derived embryos likely resulting from aberrant gene expression. To characterize three genes implicated to be important in normal preimplantation embryo development, the abundance of TRIM28, SETDB1, and TP53 was determined in in vitro fertilized (IVF), somatic cell nuclear transfer (NT), parthenogenetic, and in vivo-derived (IVV) porcine oocytes and embryos. Abundance of mRNA was quantified by real-time PCR after mRNA isolation and cDNA amplification. There was no difference in TRIM28 or SETDB1 abundance between oocytes matured in vitro versus in vivo. There was an increase of TP53 in in vitro matured oocytes. TRIM28 increased from MII to the 4-cell and blastocyst stage in IVF embryos, whereas IVV embryos have decreased TRIM28 abundance from maturation throughout development. Relative abundance of TP53 increases around the blastocyst stage in all treatment groups, but is higher in IVF embryos compared to IVV and NT embryos. SETDB1 decreases from the 2-cell to blastocyst stage in all treatments. For each gene analyzed, NT embryos of both hard to clone- and easy to clone- cell lines were more comparable to IVV embryos than IVF embryos. TRIM28, SETDB1, and TP53 are dynamically expressed in porcine oocytes and embryos. TRIM28 and TP53 are aberrantly expressed in IVF embryos in comparison to IVV and NT derived embryos. Knockdown of TRIM28 has no effect on blastocyst development or expression of SETDB1 or TP53.
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