Abstract
BackgroundNuclear 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.ResultsUsing 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.ConclusionsThese 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.
Highlights
Nuclear reprogramming reinstates totipotency or pluripotency in somatic cells by changing their gene transcription profile
The derivation of in vivo embryos, nuclear transfer (NT) embryos and donor cells with the same genetic background Embryos and cells were collected using the methods shown in Fig. 1 to enable an exact syngeneic comparison of in vivo embryos and NT embryos
NTM embryos were reconstructed with BDF1 mouse embryonic fibroblast (MEF) nuclei and the MII oocyte cytoplasm
Summary
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. Utilizing somatic cell nuclear transfer (SCNT) technology, somatic cells have been reprogrammed to totipotency and to form live cloned offspring [1]. The NT technique displays great potential in various applications, the developmental ability of NT embryos remains very poor [3]. The well-known transcriptional period ZGA is mainly regulated by maternal factors [10, 11] and epigenetic patterns [12, 13]. NT embryos appear to undergo abnormal gene regulation during this period. Certain somatic genes are constitutively expressed in 2-cell NT embryos [14].
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