Alteration of the DNA methylation status of donor cells impairs the developmental competence of porcine cloned embryos.
Alteration of the DNA methylation status of donor cells impairs the developmental competence of porcine cloned embryos.
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DOI：
10.1262/jrd.2015-048
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Liu ZH
中科院分区：
文献类型：
--
作者：
Huan YJ;Wu ZF;Zhang JG;Zhu J;Xie BT;Wang JY;Li JY;Xue BH;Kong QR;Liu ZH
体细胞核移植诱导的核重编程是一个低效率的过程，供体细胞DNA甲基化状态被认为是影响克隆效率的主要因素。本文研究了5-aza-2′-脱氧胞苷（5-aza-dC）或5-甲基-2′-脱氧胞苷-5′-三磷酸（5-甲基- dctp）调控供体细胞DNA甲基化状态在猪克隆胚胎早期发育中的作用。我们的研究结果表明，5-aza-dC或5-甲基- dctp显著降低或增加了供体细胞的整体甲基化水平，并改变了关键基因的甲基化和表达水平。然而，从这些细胞衍生的克隆胚胎的发育减少了。此外，在这些细胞衍生的克隆胚胎中，观察到伪原核形成和早期胚胎发育相关基因的转录本被破坏。综上所述，我们的研究结果表明，5-aza-dC或5-甲基- dctp对供体细胞DNA甲基化状态的改变破坏了核重编程，损害了猪克隆胚胎的发育能力。
Nuclear reprogramming induced by somatic cell nuclear transfer is an inefficient process, and donor cell DNA methylation status is thought to be a major factor affecting cloning efficiency. Here, the role of donor cell DNA methylation status regulated by 5-aza-2'-deoxycytidine (5-aza-dC) or 5-methyl-2'-deoxycytidine-5'-triphosphate (5-methyl-dCTP) in the early development of porcine cloned embryos was investigated. Our results showed that 5-aza-dC or 5-methyl-dCTP significantly reduced or increased the global methylation levels and altered the methylation and expression levels of key genes in donor cells. However, the development of cloned embryos derived from these cells was reduced. Furthermore, disrupted pseudo-pronucleus formation and transcripts of early embryo development-related genes were observed in cloned embryos derived from these cells. In conclusion, our results demonstrated that alteration of the DNA methylation status of donor cells by 5-aza-dC or 5-methyl-dCTP disrupted nuclear reprogramming and impaired the developmental competence of porcine cloned embryos.