Project description:We developed a ChIP protocol for the analysis of histone marks using less than 10,000 cells per IP, and used it to investigate the chromatin state of E11.5 mouse primordial germ cells (PGCs). A genome-wide ChIP-Seq analysis of E11.5 PGCs revealed a distribution of H3K4me3/H3K27me3 bivalent domains highly enriched for developmental regulatory genes. H3K4me3 and H3K27me3 ChIP-Seq from mouse E11.5 primordial germ cells.
Project description:We developed a ChIP protocol for the analysis of histone marks using less than 10,000 cells per IP, and used it to investigate the chromatin state of E11.5 mouse primordial germ cells (PGCs). A genome-wide ChIP-Seq analysis of E11.5 PGCs revealed a distribution of H3K4me3/H3K27me3 bivalent domains highly enriched for developmental regulatory genes.
Project description:The sexual dimorphism in mouse gonads becomes evident at around E12.5 followed by initiation of germ cell development. Previous research has been focusing on protein-coding genes. Accumulating evidence has been showing the active involvement of various ncRNAs in development. However, we are still far from having a complete picture of the RNA regulatory networks in early gonadal development. In this study, we performed sequoia complete stranded RNA-Seq using pooled total RNAs from mouse male and female gonads at E11.5, E12.5, E13.5, and E14.5, respectively. By adding poly(A) tails and unique molecular identifiers (UMI) to all stranded RNAs, we were able to quantify short- and long-stranded RNAs with or without poly(A) tails in one preparation for each time point. This is the first profiling of complete stranded RNAs during early mouse gonad development, specifically emcompassing critical stages such as sex determination and initiation of germ cell development. with this advanced sequencing technique, we aim to gain a comprehensive understanding of RNA landscape and regulatory networks involved in these critical developmental processes.
Project description:Transcriptional profiling of individual mouse embryonic (e11.5) XY gonads comparing inbred strains that are sensitive (C57BL/6J) and resistant (129S1/SvImJ) to XY sex reversal, and their reciprocal F1 hybrids. Experiment Overall Design: Two-color Agilent microarray profiles of 20 individual pairs of e11.5 XY gonads, including 5- C57BL/6J, 5- 129S1/SvImJ, 5- (B6x129S1)F1, and 5- (129S1xB6)F1 samples. Within each strain, samples were collected from multiple litters to account for potential litter biases. All samples were processed following the same protocol.
Project description:Transcriptional profiling of individual mouse embryonic (e11.5) XY gonads comparing inbred strains that are sensitive (C57BL/6J) and resistant (129S1/SvImJ) to XY sex reversal, and their reciprocal F1 hybrids.
Project description:We constructed a comparative proteome profile of female mouse fetal gonads at specific time points (11.5, 12.5, and 13.5 days post coitum), spanning a critical window for initiation of meiosis in female germ cells. We identified 3666 proteins, of which 473 were differentially expressed.
Project description:To elucidate the function of NANOS2 to regulate the transcriptome in embryonic male germ cells, we performed expression microarray analysis of the embyornic gonads of the Nanos2+/-, Nanos2-/- male and wild type female from E12.5 to E15.5. The Nanos2+/- and Nanos2-/- female gonads at E15.5 were analyzed as a negative control.
