Project description:We identified 429 potential RNA targets of DAZL in the human fetal ovary (padj<0.01), with function in synaptonemal complex formation and recombination (SYCP1, SYCP3, HORMAD1, TRIP13, TEX11), structural maintenance of chromosomes/cohesin formation and spindle assembly checkpoint (SMC1B, RAD21L, MAD2L1), and DNA repair (RAD18, RAD51, RAD54B).
Project description:The autosomal gene Dazl is a member of highly conservative DAZ family, all of which contains a RNA binding domain and expresses in germ cells. In mice, Dazl plays an essential role in germ cell development, germ cell lost in Dazl knockout mouse on pure C57BL/6 strain background. Here we use HITS-CLIP to globally characterize the genome-wide target RNA of Dazl in mouse testes. Sequencing data provides large quantities of clues to help us to get close to the truth of Dazl function. We find over 1700 RNAs are bind by Dazl mainly at 3’UTR and the genes associated with transcription, RNA splicing and reproductive cellular process are enriched significantly.
Project description:The autosomal gene Dazl is a member of highly conservative DAZ family, all of which contains a RNA binding domain and expresses in germ cells. In mice, Dazl plays an essential role in germ cell development, germ cell lost in Dazl knockout mouse on pure C57BL/6 strain background. Here we generated conditional DAZL knockout mice which allowed us to examine the roles of DAZL in postnatal spermatogenesis without affecting its early function in PGC. RNA was extracted from 10dpp wild type and Stra8-cre KO mouse testes to perform RNA-seq, StringTie software was used to assess the expression level by formula log2(FPKM+1).
Project description:Primordial germ cells (PGCs) are the embryonic precursors to egg and sperm. When removed from the embryonic gonad, PGCs can give rise to embryonic germ cell lines (EGs), pluripotent stem cells that display most of the characteristics of embryonic stem cells (ESCs) including the ability to form teratomas and to contribute to chimera formation. In mice, EG cells can be derived between E8.5 up to E12.5 of embryonic development, at which point the PGCs undergo sexual differentiation and in the male transition into unipotent gonocytes. Dazl, a germ cell-specific RNA-binding protein, is specifically expressed in developing PGCs and is required for proper germ cell development. Dazl knockout mice are infertile, but the molecular mechanisms underlying this phenotype are still unknown. Here we demonstrate that Dazl localizes in granular structures in mammalian PGCs but not in ESCs. We demonstrate Dazl plays a central role in a large mRNA/protein interactive network that includes members of Fragile-X family RNA-binding proteins. We demonstrate that Dazl and Fxr1 play a central role in these granules and directly regulate the translation of specific core pluripotency factors, including Sox2 and Suz12. Global gene expression changes following Dazl knockdown in in vitro primordial germ cells. In vitro primordial germ cells carrying control and Dazl knockdown shRNAs were generated from Oct4-GFP ES cells and isolated by FACS analysis. The global gene expression profiles were analyzed by Agilent Mouse Whole Genome 4X44K one-color microarrays.
Project description:Dazl (deleted in azoospermia like) is a member of the DAZ family of germ cell-restricted RNA binding proteins required for gametogenesis from worm to human. The direct RNA targets and functions of these essential proteins are poorly understood. Here, we generated high-resolution, transcriptome-wide maps of Dazl-RNA interactions in mouse testes. These maps provide important insights into the mechanism of Dazl recruitment to mRNA and reveal Dazl binding to thousands of mRNAs predominantly through sequence-specific interactions near the polyA tail. Using transgenic mice and fluorescence activated cell sorting (FACS), we isolated DAZL knockout germ cells and used RNA-Seq to identify mRNAs sensitive to DAZL-ablation. Intersecting the RNA-Seq and Dazl-RNA interaction datasets revealed that Dazl enhances expression of a subset of directly-bound transcripts, namely mRNAs for a network of essential cell cycle regulatory genes. Collectively, our integrative analysis delineates a Dazl-dependent post-transcriptional gene regulatory program essential for mammalian germ cell maintenance.
Project description:We have identified a Golgi- and microtubule-rich vesicular structure, "fusome" which resembles the Drosophila fusome and accumulates asymmetrically during germ cell cyst divisions starting at E11.5. Genes associated with fusome become enriched in presumptive oocytes during cyst divisions and early meiosis, suggesting a role in oocyte rejuvenation. To investigate this further, we performed single-cell RNA sequencing (scRNA-seq) of mouse fetal germ cells at E10.5, E11.5, and E15.5, including developmental stages not previously covered in earlier datasets (GSE136441), thereby generating a composite dataset of E10.5-P5 germ cells. Additionally, we analyzed Dazl homozygous and heterozygous mutant ovarian germ cells at E11.5 and E12.5 to study the impact of Dazl loss on cyst formation and fusome-associated gene expression dynamics. This dataset provides a comprehensive resource to explore transcriptional changes across key stages of fetal ovarian development and in the context of Dazl function.
Project description:Dazl (deleted in azoospermia like) is a member of the DAZ family of germ cell-restricted RNA binding proteins required for gametogenesis from worm to human. The direct RNA targets and functions of these essential proteins are poorly understood. Here, we generated high-resolution, transcriptome-wide maps of Dazl-RNA interactions in mouse testes. These maps provide important insights into the mechanism of Dazl recruitment to mRNA and reveal Dazl binding to thousands of mRNAs predominantly through sequence-specific interactions near the polyA tail. Using transgenic mice and fluorescence activated cell sorting (FACS), we isolated DAZL knockout germ cells and used RNA-Seq to identify mRNAs sensitive to DAZL-ablation. Intersecting the RNA-Seq and Dazl-RNA interaction datasets revealed that Dazl enhances expression of a subset of directly-bound transcripts, namely mRNAs for a network of essential cell cycle regulatory genes. Collectively, our integrative analysis delineates a Dazl-dependent post-transcriptional gene regulatory program essential for mammalian germ cell maintenance.