Single Cell RNA Sequencing Reveals The Difference Of Transcriptomes In SCNT Embryos Drived From Different Somatic Cell Type

In the field of nuclear transplantation on the most famous is the Gurdon and Wilmut two,since Wilmut successfully created the world's first cloned mammal cloned sheep "Dolly",to study the nuclear transplantation technique in mammals greatly promote.Although so far there are many species of mammals were successfully cloned,but from the first cloned animal reports from now has nearly 59 years of history,the current nuclear transfer efficiency in higher organisms is still very low,with a birth rate of about 1% to 3%.For nuclear transfer embryonic development of low scientific problems,but also because of nuclear heat transfer technology has become a hot research category.The focus of the study was then focused on the mechanism of early embryonic development.One of the most obvious phenomena of cloned embryos is the early cleavage arrest,which is consistent with the activation of the embryonic genome.It occurs in the 2-cell stage in mice,and there are differences in other species.The development of cloned embryos based on the laboratory selection of B6D2F1 hybrid mouse in vivo normal embryos,cumulus cell nuclear transfer embryos and MEF embryos three group 2-cell embryos,as research material,using the single cell sequencing around 2-cell embryo development block analysis of transcriptome data.Through the analysis of the transcriptome data,some important genes related to the development of 2-cell embryo were screened out.The use of clustering,thermography and GO annotation analysis methods such as analysis of 2-cell during the period of transcriptome data of different embryos,the sequencing results reveal the gene expression differences were screened out in this paper,nuclear transplantation embryos significantly down regulated the expression of these genes;differential gene expression in three groups of embryos were screened out in nuclear transplantation compared with the normal in vivo embryo embryo,a number of genes expressed more apparent differences in the transcriptome data;genetic related genes,OSKM transcription factors and cell cycle related genes analysis showed normal embryos and in vivo nuclear transplantation embryos between the gene expression level difference;analysis of new transcription transcriptome sequencing of the gene shows,these new transcription of this gene in vivo normal embryo is generally highly active,and in nuclear transplantation embryos lower expression,and in the embryo is not the most active in cumulus cell nuclear transplantation.To sum up,there are significant differences in the transcription of 2-cell embryos.A comparative analysis of difference analysis in this thesis,which is a small part of gene expression has been reported related to the 2-cell block gene,many genes in embryonic development,although not a 2-cell function,it may be inferred that this gene is also closely related with the embryonic 2-cell block.It is a way to improve the rate of early embryonic development to correct the abnormal expression of genes in the 2-cell period with low or over expression.The results of the transcriptome analysis in this study not only reveal the difference between nuclear transfer embryos and normal embryonic transcripts in vivo,but also provide valuable clues and guidance for studying the 2-cell phase-related studies of nuclear transfer embryos.