Project description:Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and Larp7 Knock-out Staged Male Germ Cells Transcriptomes
Project description:Mass spectrometry data of RNA interactome capture (RIC) assay for STA-PUT purified mouse primary male germ cells, including spermatogonia (SG), pachytene spermatocyte (pSC) and round spermatid (rST)
Project description:We seprated the primodial germ cells using MACS from E12.5 stages in WT and Rif1-knock out mice, and using modified Smart-seq2 methods to get 1,000 cells library.
Project description:Analysis the difference in the transcriptomes (expression and alternative splicing) of either spermatocytes or round spermatids between Larp7 conditional knock-out and control mice
Project description:The mouse zinc finger genes Zfy1 and Zfy2 are essential for male fertility. Recently, we produced Zfy1 knock-out (KO), Zfy2 KO, and Zfy1/2 double-knock-out (Zfy DKO) mice, and found that Zfy DKO males were infertile. The mechanism by which ZFY contributes to reproduction remains unknown but based on predicted protein sequence and in vitro assays we hypothesize that it controls expression of genes essential for spermatogenesis. To identify which genes ZFY regulates, we performed comparative transcriptome analysis of sorted male germ cells at three different spermatogenesis stages from three Zfy KO models and control wild-type males. Significantly altered germ cell transcriptomes were identified with Zfy2 KO and Zfy DKO. Analyses of differentially expressed genes supported that Zfy loss altered spermatogenesis, DNA packaging/chromatin organization, and apoptosis pathways. Alternative splicing was deregulated in Zfy KO models, affecting sperm function and chromatin regulation pathways. In support of in-silico findings, Zfy DKO males were shown to have impaired sperm chromatin organization, functional sperm deficiencies, and increased germ cell apoptosis. ZFY regulation of apoptotic pathways was demonstrated also in transfected human cells. We conclude that Zfy is a critical regulator of meiosis and spermiogenesis in addition to its previously described function as a cell-cycle regulator.