Project description:The present investigation was to identify transcriptomic changes of control and Dnd1-cKO PGCs at E12.5 and E13.5 by microarray comparison. We identified 1470 upregulated and 924 downregulated genes in Dnd1-cKO PGCs.
Project description:The present investigation was to identify transcriptomic changes of control and Dnd1-cKO PGCs at E11.5 by RNA-seq analysis. We identified 181 upregulated and 141 downregulated genes in Dnd1-cKO PGCs.
Project description:Specification of germ cell fate establishes the germline development during early embryogenesis, yet the underlying mechanisms remain largely unknown in humans. Here we focus on the functional roles of the RNA-binding protein (RBP) DND1 in human germline specification. We deleted the whole genomic region of DND1 in human embryonic stem cells (hESCs), based on which we generated human primordial germ cell-like cells (hPGCLCs). Interestingly, we discovered an increased percentage of hPGCLCs induced from DND1 deleted hESCs, suggesting that DND1 may restrict the specification of human germ cell lineage. Mechanistic investigation reveals that DND1 forms a complex with another RBP NANOS3, in which DND1 facilitates the binding of NANOS3 to their target mRNAs. Furthermore, by analyzing the mRNAs bound by DND1 and NANOS3, we identified SOX4 mRNAs as the key downstream factor for DND1 and NANOS3 complex to restrict the induction of hPGCLCs. Interestingly, DND1 and NANOS3 function in processing bodies (P-bodies) to repress the translation of SOX4 mRNAs, where NANOS3 bridges the interaction between DND1 and the translational repressor 4E-T. Altogether, these findings identify the RBPs DND1 and NANOS3 as “break system” to restrict the entry of germ cell fate in humans.
Project description:Purpose: The purpose of this study is to compare the transcriptome expression profiles of E12.5 and E13.5 Osr2RFP/- and Osr2RFP/+ palatal mesenchyme by using RNA-seq analysis. Methods: Osr2RFP/+ male mice were crossed with Osr2+/- female mice. The embryos were harvested at E12.5 and E13.5. The pair of palatal shelves were dissected from each Osr2-RFP positive embryo. The RFP+ palatal mesenchyme cells were isolated by using fluorescence-activated cell sorting (FACS). RNA-seq analysis was carried out using the FACS-isolated palatal mesenchyme from Osr2RFP/- and Osr2RFP/+ embryos, respectively.
Project description:Mouse embryonic skin samples were collected at E13.5 or E14.5. For each condition, dorsolateral skin from littermate control and mutant embryos was dissected and dissociated for single-cell RNA-seq. Conditions include constitutive dermal β-catenin activation (Actβcat, E13.5), SHH overexpression (SHH OE, E13.5), and Gli3 conditional knockout (Gli3 cKO, E13.5), each with paired littermate controls. An additional E14.5 wildtype replicate was included.
Project description:Analysis of our transcriptome and RNA immunoprecipitation experiments indicate DND1 acts as a positive regulator of chromatin modifiers in male germ cells, linking RNA binding proteins and epigenetic regulation.
Project description:During embryonic development, large-scale reprogramming occurs in primordial germ cells (PGCs) at the level of both DNA demethylation and histone modifications. While much is known about epigenetic changes, it still remains unclear how this chromatin state impacts the transcriptome and development of the germline. We have used cell-number normalised (CNN) RNA-seq to document the striking upregulation of the germ cell transcriptome at E13.5 compared to neighboring somatic cells of the embryonic gonad. We provide a novel, genome-wide analysis of hypertranscription during development, documenting the global amplification of the majority of the E13.5 PGC transcriptome. Upregulated transcripts in PGCs include those for ribosome biogenesis, translation and chromatin remodelling, as well as all transposable element families. These data reveal the significant hyperactivity of the germ cell transcriptome.