Project description:In human embryos, major zygotic genome activation (ZGA) initiates at the eight-cell (8C) stage. Abnormal ZGA leads to developmental defects and even contributes to the failure of human blastocyst formation or implantation. However, little is known about the mechanisms regulating human ZGA, because of many challenges and constraints on research on human embryos. Here we report one type of rare 8C-like cells (8CLCs) that are discovered specifically from human preimplantation epiblast-like stem cells (prEpiSCs) and naïve embryonic stem cells. 8CLCs express a list of human ZGA genes and their transcriptome closely resemble that of the human 8C embryo. An 8C-specific reporter is further developed to isolate 8CLCs from prEpiSCs and optimize the chemical-based culture condition that increases and maintains the 8CLC population. Functionally, 8CLCs can spontaneously form blastocyst-like structures. The discovery and maintenance of 8CLCs provides an opportunity to model human ZGA and recapitulate early human development.
Project description:In human embryos, major zygotic genome activation (ZGA) initiates at the eight-cell (8C) stage. Abnormal ZGA leads to developmental defects and even contributes to the failure of human blastocyst formation or implantation. However, little is known about the mechanisms regulating human ZGA, because of many challenges and constraints on research on human embryos. Here we report one type of rare 8C-like cells (8CLCs) that are discovered specifically from human preimplantation epiblast-like stem cells (prEpiSCs) and naïve embryonic stem cells. 8CLCs express a list of human ZGA genes and their transcriptome closely resemble that of the human 8C embryo. An 8C-specific reporter is further developed to isolate 8CLCs from prEpiSCs and optimize the chemical-based culture condition that increases and maintains the 8CLC population. Functionally, 8CLCs can spontaneously form blastocyst-like structures. The discovery and maintenance of 8CLCs provides an opportunity to model human ZGA and recapitulate early human development.
Project description:In human embryos, major zygotic genome activation (ZGA) initiates at the eight-cell (8C) stage. Abnormal ZGA leads to developmental defects and even contributes to the failure of human blastocyst formation or implantation. However, little is known about the mechanisms regulating human ZGA, because of many challenges and constraints on research on human embryos. Here we report one type of rare 8C-like cells (8CLCs) that are discovered specifically from human preimplantation epiblast-like stem cells (prEpiSCs) and naïve embryonic stem cells. 8CLCs express a list of human ZGA genes and their transcriptome closely resemble that of the human 8C embryo. An 8C-specific reporter is further developed to isolate 8CLCs from prEpiSCs and optimize the chemical-based culture condition that increases and maintains the 8CLC population. Functionally, 8CLCs can spontaneously form blastocyst-like structures. The discovery and maintenance of 8CLCs provides an opportunity to model human ZGA and recapitulate early human development.
Project description:After fertilization, zygotic genome activation (ZGA) enables the conversion of two terminally differentiated gametes to a totipotent embryo. Zygotes further give rise to the pluripotent embryonic lineages and extraembryonic trophectoderm after the first lineage commitment. While much is learned for pluripotency regulation, how ZGA is connected to the pluripotency commitment in early embryos remains unclear. Here, we investigated the role of nuclear receptor (NR) family TFs in mouse pre-implantation embryos, whose motifs are highly enriched in accessible chromatin at the 2-cell (2C) to 8-cell (8C) stages. We found NR5A2 is required for the early development, as both knockdown and knockout of Nr5a2 led to morula arrest. 4-8C activated genes (mid-preimplantation activation), including key pluripotency marker genes (i.e. Nanog, Pou5f1, and Tdgf1) and trophectoderm genes (i.e. Klf5, Elf3, and Gata3). Genome-wide chromatin binding and RNA-seq analyses showed NR5A2 bound and regulated the 4-8C genes, including both ICM and TE genes in 2C and 8C embryos , indicating its roles in bipotency program. Interestingly, NR5A2 occupied sites predominantly reside in accessible B1 elements where its motif is embedded at the 2-8C stage. Taken together, these data demonstrate the role of NR5A2 as a key regulator that bridges ZGA to lineage segregation.
Project description:Recapitulating early mammalian kidney development in vitro by priming and differentiating mouse embryonic stem cells as a monolayer