Transcriptome Analyses Reveal Effects of Vitamin C-Treated Donor Cells on Cloned Bovine Embryo Development.
Transcriptome Analyses Reveal Effects of Vitamin C-Treated Donor Cells on Cloned Bovine Embryo Development.
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转录组分析揭示维生素 C 处理的供体细胞对克隆牛胚胎发育的影响
DOI：
10.3390/ijms20112628
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
郭泽坤
中科院分区：
文献类型：
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作者：
郭泽坤
体细胞核移植（SCNT）是一种非常强大的技术，用于生产基因相同或修饰的动物。然而，哺乳动物的克隆效率仍然很低。在这项研究中，我们的目的是探讨维生素C（Vc）处理的供体细胞对克隆胚胎的影响。结果，Vc处理放松了供体细胞的染色质，改善了克隆胚胎的发育。采用RNA测序技术研究了早期胚胎中转录谱的变化。我们发现，Vc处理增加了参与细胞-基质粘附连接的基因的表达。基因本体（GO）分析表明，Vc处理促进了自噬的激活，这是缺乏克隆的两细胞胚胎。雷帕霉素是一种有效的自噬激活剂，可增加克隆囊胚的形成（36.0% vs.25.6%，p < 0.05）。Vc处理恢复了克隆胚中一些编码基因和长链非编码RNA的异常表达，包括锌指蛋白641（ZNF 641）。ZNF 641通过mRNA显微注射补偿提高了克隆胚胎的发育潜力。此外，Vc处理挽救了克隆的两细胞胚胎中一些缺陷的RNA编辑位点。总的来说，VC处理的供体细胞通过影响胚胎转录来改善克隆胚胎的发育。本研究为今后促进SCNT胚胎重编程过程的研究提供了有用的资源。
Somatic cell nuclear transfer (SCNT) is a very powerful technique used to produce genetically identical or modified animals. However, the cloning efficiency in mammals remains low. In this study, we aimed to explore the effects of vitamin C (Vc)-treated donor cells on cloned embryos. As a result, Vc treatment relaxed the chromatin of donor cells and improved cloned embryo development. RNA sequencing was adopted to investigate the changes in the transcriptional profiles in early embryos. We found that Vc treatment increased the expression of genes involved in the cell–substrate adherens junction. Gene ontology (GO) analysis revealed that Vc treatment facilitated the activation of autophagy, which was deficient in cloned two-cell embryos. Rapamycin, an effective autophagy activator, increased the formation of cloned blastocysts (36.0% vs. 25.6%, p < 0.05). Abnormal expression of some coding genes and long non-coding RNAs in cloned embryos was restored by Vc treatment, including the zinc-finger protein 641 (ZNF641). ZNF641 compensation by means of mRNA microinjection improved the developmental potential of cloned embryos. Moreover, Vc treatment rescued some deficient RNA-editing sites in cloned two-cell embryos. Collectively, Vc-treated donor cells improved the development of the cloned embryo by affecting embryonic transcription. This study provided useful resources for future work to promote the reprogramming process in SCNT embryos.