Project description:Our data delineated bifurcated binding and regulation of ZFP352 towards two distinct retrotransposons, MT2_Mm and SINE_B1/Alu,further ChIP-seq experiments validated that over-expression of Dux displaced ZFP352 from SINE_B1/Alu and DUX-independent ZFP352 binding sites, but promoted ZFP352 binding onto MT2_Mm and DUX_ZFP352 co-bound sites
Project description:Previous study has determined Dux regulates 2CLC trancriptome via binding and activating MERVL,but the mechanism about the change happned during the 2C-exit process are rarely determined.In this study,we try to illustrate how ZFP352 binds genome location to regulate its downstream trancriptome and 2C-exit process.And make some conclusion and hypothesis about ZFP352 co-binding and interact with other trancription factor:Dux,etc to regulate 2C-exit.
Project description:Our data delineated bifurcated binding and regulation of ZFP352 towards two distinct retrotransposons, MT2_Mm and SINE_B1/Alu,and whether ZFP352 can regulate mouse genome openness remains unknown,here we conduct ATAC-seq of Zfp352 over expression sample to detect ZFP352's role in regulating chromatin openness.
Project description:Mouse ES cell population contains a minor sub-population expressing the genes that are specifically expressed in 2-cell stage embryos. This sub-population, consists of the 2-cell-gene labelled cells (2CLCs), is generated by the transient activation of the 2-cell specific genes initiated by the master regulator Dux. However, the mechanism regulating the transient expression remains largely unclear. Here we reported a novel function of Zfp352, one of the 2-cell specific genes, in the regulation of the 2CLC sub-population. Zfp352 encodes zinc-finger transcription factor belonging to the Klf family. Zfp352 is transiently activated by Dux after the activation of Zscan4c and expresses in a subset of 2CLC subpopulation. Interestingly, in the reporter assay, the transcriptional activation of Zscan4c by Dux is strongly repressed by the co-expression of Zfp352. However, knockout of Zfp352 resulted in the repression of a subset of the 2-cell specific genes. These data suggest the dual roles of Zfp352 in the regulation of the transient activation of the 2-cell specific genes.
Project description:Prevous study has discoverd 2CLC trancriptome and MERVL can be up-regulated by Dux over expression,but rarely has been found about the whole procedure of Dux-induced 2C-entry and exit process and whether this system can reflect the natural 2-cell embryo development,so we we defined different time point transcription after Dux over expression,and try to establish a link between gene expression profile between gene dynamics in different Dux over expression time point and in early mouse embryo development
Project description:Acquisition of new stem cell fates relies on the dissolution of the prior regulatory network sustaining the existing cell fates. Currently, extensive insights have been revealed for the totipotency regulatory network around the zygotic genome activation (ZGA) period. However, how the dissolution of the totipotency network is triggered to ensure the timely embryonic development following ZGA is largely unknown. In this study, we identified the unexpected role of a highly expressed 2C embryo specific transcription factor, ZFP352, in facilitating the dissolution of the totipotency network. We found that ZFP352 has bifurcated binding and regulation towards two different retrotransposon sub-families. ZFP352 coordinates with DUX to bind and regulate the 2C specific MT2_Mm sub-family. On the other hand, without DUX, ZFP352 switched affinity to bind extensively onto SINE_B1/Alu sub-family. This leads to the activation of later developmental programs like ubiquitination pathways, to facilitate the dissolution of the 2C state. Correspondingly, depleting ZFP352 in mouse embryos impaired the 2C to morula transition process. Thus, through differential regulation of MT2_Mm and SINE_B1/Alu, ZFP352 can trigger spontaneous dissolution of the totipotency network. Our study highlights the importance of different retrotransposons sub-families in facilitating the timely and programmed cell fates transition during early embryogenesis.
Project description:We report both DUX4 and Dux toxicity depend upon their ability to bind DNA and activate transcription. Chromatin immunoprecipitation of V5 epitope tagged human DUX4 and mouse Dux was performed in human myoblasts was analyzed using ChIP-Seq to identify their subsequent binding sites. We found that DUX4 and Dux bind 4-8% of identical sequences, while majority of the binding sites are unique to either DUX4 or Dux. Although small, this overlap could be due to their conserved abilioty to regualte primordial pathways that were essential for life and therefore maintained in both proteins despite their separate evolutionary paths. We performed ChIP-Seq analysis of human myoblasts transfected with plasmids encoding either epitope tagged human DUX4 (1 sample) and mouse Dux (1 sample). Illumina sequencing libraries were prepared from the ChIP and Input DNA, then resulting DNA libraries were quantified and sequenced and aligned to the human genome (hg19).
Project description:To explore the function of ZFP352 during mouse early embryonic development, siRNA targeting Zfp352 was injected into the zygotes and the effect of Zfp352 depletion on development was followed.