Project description:To gain more insights about the role of Hbxip in mESC, we performed RNA-Seq for WT and Hbxip-KO E14 ESCs in pluripotent condition and during differentiation. The goals of this study are to compare the transcriptome changes upon Hbxip knockout in pluripotent condition and during differentiation.
Project description:We analyzed the genome-wide binding of Sox2 and POU factor partner factors, Oct4 in ESCs (using published datasets PMID:18692474 and GSM307137, GSM307154, GSM307155) and Brn2 in NPCs. We found that Sox2 and Oct4 co-occupied a large subset of promoters and enhancers in ESCs, but that Sox2 and Brn2 co-occupy predominantly enhancers. Further, we overexpressed Brn2 in differentiating ESCs and showed that ectopic Brn2 recruited Sox2 to NPC-specific targets, resulting in skewed differentiation towards the neural lineage. Examination of transcription factor binding in ESCs, NPCs, and differentiating ESCs by ChIP-Seq.
Project description:We analyzed the genome-wide binding of Sox2 and POU factor partner factors, Oct4 in ESCs (using published datasets PMID:18692474 and GSM307137, GSM307154, GSM307155) and Brn2 in NPCs. We found that Sox2 and Oct4 co-occupied a large subset of promoters and enhancers in ESCs, but that Sox2 and Brn2 co-occupy predominantly enhancers. Further, we overexpressed Brn2 in differentiating ESCs and showed that ectopic Brn2 recruited Sox2 to NPC-specific targets, resulting in skewed differentiation towards the neural lineage.
Project description:Mouse embryonic stem cells (ESCs) sporadically express preimplantation two-cell-stage (2C) transcripts, including MERVL endogenous retrovirus and Zscan4 cluster genes. Such 2C-like cells (2CLCs) can contribute to both embryonic and extraembryonic tissues when reintroduced into early embryos. We examined global nucleosome occupancy and gene expression in 2CLCs and identified miR-344 as the noncoding molecule that positively controls 2CLC potency. We found that activation of endogenous MERVL or miR-344-2 alone is sufficient to induce 2CLCs with induction of 2C genes and an expanded potency. Mechanistically, miR-344 is activated by the 2C-state driver DUX and post-transcriptionally represses ZMYM2 and LSD1, which recruit the HDAC corepressor to MERVL LTR for transcriptional repression. Consistently, zygotic depletion of Zmym2 compromises the totipotency-to-pluripotency transition during early development. Our studies establish the novel DUX->miR-344--|Zmym2/Lsd1 axis that controls MERVL for expanded stem cell potency.
Project description:The interaction between SUMO and ZMYM2 is mediated through specific motifs found in ZMYM2 that are called SIMs (SUMO interacting motifs). We found/characterised three SIMs in ZMYM2 and in vitro experiments showed that mutation in these residues prevent the interaction with SUMO. Microarray transcription profiling was done to study the effects of disrupting the multiSUMO binding activity of ZMYM2, assessing the differences in gene expression in cells expressing wild-type or SIM2 mutant version of ZMYM2.
Project description:We performed bulk mRNA sequencing of either control or Zfp266 KO mouse embryonic fibroblasts (MEFs), intermediate reprogramming populations, iPSCs and ESCs. We reveal enhanced and early upregulation of pluripotent and ESC associated genes in Zfp266 KO reprogramming populations.