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of Biology-Oriented Science and Technology, Kindai Universityhttps://www.ebi.ac.uk/ena/browser/view/PRJNA681570We have developed a nuclear transfer (NT) system in which somatic nuclei are transplanted into mouse embryos arrested at the 4-cell stage. The transplanted somatic nuclei show swelling and epigenetic reprogramming towards 4-cell-like nuclei. To assess genome-wide transcriptional reprogramming of the injected nuclei, the newly transcribed genes in NT embryos were examined by RNA-seq analyses. As a control, NT was also performed using mouse embryos at the 2-cell stage. Overall design: Mouse C2C12 cells or primary Oryx dammah cells were fused with mouse 4-cell embryos arrested at G2/M phase with 0.1 µg/ml demecolcine. The reconstructed NT embryos were incubated in mouse embryo culture medium containing 0.1 µg/ml demecolcine with or without alpha-amanitin. After 24 hours of culture, the NT embryos were subjected to RNA-seq analyses to detect newly transcribed genes in a RNA polymerase II-dependent manner by comparing embryos without alpha-amanitin and those with alpha-amanitin. Furthermore, to examine the effect of DNA replication, nuclear transfer of C2C12 cells to 4-cell embryos was performed before and after DNA replication at the 4-cell stage (40 hours post insemination [hpi] and 46 hpi, respecitively, followed by 24 hours of culture). Then, transcriptomes of these embryos with or without DNA replication were compared. Reprogramming ability of 2-cell embryos was also examined by transplanting C2C12 cells to early (21 hpi) or late 2-cell embryos 30 hpi).xref:PubMed:35463476xref:PubMed:34849463ENAdTAF[[II]]230, TAF[[II]]250, d230, Laboratory, Mus domesticus, mouse, TAF200, l(3)84Ab, dTAFII250, BG:DS00004.13, TAFII-250, TAF250/230, EfW1, House Mouse, Cell, mice C57BL/6xCBA/CaJ hybrid, Mus muscaris, dTAF230, dmTAF[[II]]230, TAFII250, House, Mus, dmTAF1, Taf230, p230, Mus musculus domesticus, TAF[[II]]250/230, TFIID, Mice, TAF250, Taf[[II]]250, Mus musculus, Taf200, dTAF[[II]]250, TAF[[II]]230, TFIID TAF250, cel, Swiss, cell, mice, Swiss Mouse, House Mice, Swiss Mice, Taf1p, TAF[II]250, CG17603, TAF[[II]], domesticus, Laboratory Mice, dTAF250, DmelCG17603, TAF1., Taf250, SR3-5, Mouse, house mouse, TAF, Laboratory Mouse, TAF230dTAF[[II]]230, TAF[[II]]250, d230, Laboratory, Mus domesticus, mouse, TAF200, l(3)84Ab, dTAFII250, BG:DS00004.13, TAFII-250, TAF250/230, EfW1, House Mouse, Cell, mice C57BL/6xCBA/CaJ hybrid, Mus muscaris, dTAF230, dmTAF[[II]]230, TAFII250, House, Mus, dmTAF1, Taf230, p230, Mus musculus domesticus, TAF[[II]]250/230, TFIID, Mice, TAF250, Taf[[II]]250, Mus musculus, Taf200, dTAF[[II]]250, TAF[[II]]230, TFIID TAF250, cel, Swiss, cell, mice, Swiss Mouse, House Mice, Swiss Mice, Taf1p, TAF[II]250, CG17603, TAF[[II]], domesticus, Laboratory Mice, dTAF250, DmelCG17603, TAF1., Taf250, SR3-5, Mouse, house mouse, TAF, Laboratory Mouse, TAF230falseTranscriptional reprogramming of somatic cells after nuclear transfer to mouse 4-cell embryosTranscriptional reprogramming of somatic cells after nuclear transfer to mouse 4-cell embryos2023-05-192021-11-05PRJNA681570GSE1623453546347634849463