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Transcriptional defects and reprogramming barriers in somatic cell nuclear reprogramming as revealed by single-embryo RNA sequencing

机译:单胚胎RNA测序揭示的体细胞核重新编程中的转录缺陷和重编程障碍

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摘要

Abstract Background Nuclear reprogramming reinstates totipotency or pluripotency in somatic cells by changing their gene transcription profile. This technology is widely used in medicine, animal husbandry and other industries. However, certain deficiencies severely restrict the applications of this technology. Results Using single-embryo RNA-seq, our study provides complete transcriptome blueprints of embryos generated by cumulus cell (CC) donor nuclear transfer (NT), embryos generated by mouse embryonic fibroblast (MEF) donor NT and in vivo embryos at each stage (zygote, 2-cell, 4-cell, 8-cell, morula, and blastocyst). According to the results from further analyses, NT embryos exhibit RNA processing and translation initiation defects during the zygotic genome activation (ZGA) period, and protein kinase activity and protein phosphorylation are defective during blastocyst formation. Two thousand three constant genes are not able to be reprogrammed in CCs and MEFs. Among these constant genes, 136 genes are continuously mis-transcribed throughout all developmental stages. These 136 differential genes may be reprogramming barrier genes (RBGs) and more studies are needed to identify. Conclusions These embryonic transcriptome blueprints provide new data for further mechanistic studies of somatic nuclear reprogramming. These findings may improve the efficiency of somatic cell nuclear transfer.
机译:摘要背景核编程通过改变其基因转录简介,恢复整体细胞中的全能或多能性。该技术广泛应用于医学,畜牧业等行业。然而,某些缺陷严重限制了这项技术的应用。结果采用单胚胎RNA-SEQ,我们的研究提供了由积云细胞(CC)供体核转移(NT)产生的胚胎产生的完整转录组蓝图,由小鼠胚胎成纤维细胞(MEF)供体NT和每个阶段的体内胚胎产生的胚胎( Zygote,2细胞,4细胞,8细胞,森拉和胚泡)。根据进一步分析的结果,NT胚胎表现出在基因组织激活(ZGA)期间的RNA加工和翻译起始缺陷,并且在囊胚形成期间蛋白激酶活性和蛋白质磷酸化是缺陷的。两千三个恒定基因无法在CCS和MEF中重新编程。在这些常数基因中,在所有发育阶段中连续错误转录136个基因。这136个差异基因可以是重编程的屏障基因(RBG),并且需要更多的研究来鉴定。结论这些胚胎转录组蓝图为各体核重编程的进一步机械研究提供了新数据。这些发现可以提高体细胞核转移的效率。

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