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克隆和体外受精猪胚胎中 DNA 甲基化模式的特定位置分析。

Locus-specific analysis of DNA methylation patterns in cloned and in vitro fertilized porcine embryos.

机构信息

Fujian Provincial Key Laboratory for the Prevention and Control of Animal Infectious Diseases and Biotechnology, College of Life Sciences, Longyan University, Longyan 364012, P. R. China.

Provincial Key Laboratory for Developmental Biology and Neurosciences, College of Life Sciences, Key Laboratory of Opto-Electronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou 350007, P. R. China.

出版信息

J Reprod Dev. 2020 Dec 22;66(6):505-514. doi: 10.1262/jrd.2019-076. Epub 2020 Sep 8.

Abstract

Porcine somatic cell nuclear transfer (SCNT) is currently inefficient, as 1-3.95% of reconstructed embryos survive to term; inadequate or erroneous epigenetic reprogramming of the specialized donor somatic nucleus could be a primary reason. Therefore, a locus-specific analysis of DNA methylation dynamics in embryogenesis and the DNA methylation status of gametes and donor cells used for SCNT were conducted in the following developmentally important gene loci: POU5F1, NANOG, SOX2, H19, IGF2, IGF2R, XIST; and the retrotransposon LINE-1. There were significant epigenetic differences between the gametes and the somatic donor cells. Three gamete-specific differentially methylated regions (DMRs) in POU5F1, XIST, and LINE-1 were identified. A delayed demethylation process at POU5F1 and LINE-1 loci occurred after three successive cleavages, compared to the in vitro fertilized (IVF) embryos. Although cloned embryos could undergo de-methylation and re-methylation dynamics at the DMRs of imprinted genes (H19, IGF2R, and XIST), the re-methylation process was compromised, unlike in fertilized embryos. LINE-1 loci are widely dispersed across the whole genome, and LINE-1 DMR might be a potential porcine nuclear reprogramming epi-marker. Data from observations in our present and previous studies, and two published articles were pooled to produce a schematic diagram of locus-specific, DNA methylation dynamics of cloned and IVF embryos during porcine early embryogenesis. This also indicated aberrant DNA methylation reprogramming events, including inadequate DNA demethylation and insufficient re-methylation in cloned embryos. Further research should focus on mechanisms underlying demethylation during the early cleavage of embryos and de novo DNA methylation at the blastocyst stage.

摘要

猪体细胞核移植(SCNT)的效率目前较低,只有 1-3.95%的重构胚胎能存活到足月;供体细胞特化核的表观遗传重编程不足或错误可能是主要原因。因此,在以下重要的发育基因座中,对胚胎发生过程中的 DNA 甲基化动态和用于 SCNT 的配子和供体细胞的 DNA 甲基化状态进行了局灶性分析:POU5F1、NANOG、SOX2、H19、IGF2、IGF2R、XIST 和反转录转座子 LINE-1。配子和体细胞核供体细胞之间存在明显的表观遗传差异。在 POU5F1、XIST 和 LINE-1 中鉴定出 3 个配子特异性差异甲基化区域(DMR)。与体外受精(IVF)胚胎相比,POU5F1 和 LINE-1 基因座的去甲基化过程在连续三次分裂后发生延迟。尽管克隆胚胎可以在印迹基因(H19、IGF2R 和 XIST)的 DMR 上经历去甲基化和再甲基化动态,但再甲基化过程受到了损害,与受精胚胎不同。LINE-1 基因座广泛分布于整个基因组,LINE-1 DMR 可能是猪核重编程的潜在表观遗传标记。来自我们目前和以前的研究以及两篇已发表文章的数据被汇集在一起,以生成猪早期胚胎发生过程中克隆和 IVF 胚胎的局灶性、DNA 甲基化动态的示意图。这也表明了异常的 DNA 甲基化重编程事件,包括克隆胚胎中 DNA 去甲基化不足和再甲基化不足。进一步的研究应集中在胚胎早期分裂过程中去甲基化和囊胚阶段从头 DNA 甲基化的机制上。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f138/7768172/05ccaf815410/jrd-66-505-g001.jpg

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