Project description:Loss of Mgat1 in spermatogonia was investigated in germ cells from 23 day males. Gene expression changes induced by deletion of Mgat1 were determined using the Affymetrix GeneChip Mouse Gene 2.0 ST Array. We include the expression data obtained from three replicate aliquots of pooled testis total RNA obtained from six control or six conditional knockout Mgat1 testis germ cells from 23 dpp males, Three replicates of the respective control and Mgat1 cKO pools were used for microarray analysis using Affymetrix GeneChip Whole Transcript Plus Reagent Kit. We identified differential expressed genes with a significance level of ANOVA and FDR p < 0.05 and an absolute fold-change (linear) less than -2, and greater than 2.0.
Project description:Celf1 germline or conditional deletion mouse mutants exhibit fully penetrant lens defects including cataract. To gain insight into gene expression changes underlying these lens defects, microarray comparison was performed for lenses obtained from control and Celf1 conditional deletion mutant mice.
Project description:We utilized the Illumina MouseRef-8 gene expression technology to quantify differential gene expression between wildtype mice and mice with Osterix driven Cre conditional knockout of Hdac3 (Hdac3-CKO). We compared the RNA extracted from calvaria from 8 wildtype and 8 conditional knockout litter matched mice using two separate Illumina MouseRef-8 chips. Mice with exon 7 of Hdac3 flanked by loxP sites were crossed with mice expressing Cre driven by the Osterix promoter. RNA from 5 day old mouse calvarial explants (digested for 20 minutes with collagenase) was purified using TRIzol according to the manufacturerâs protocol (Invitrogen) and reverse transcribed using Qiagenâs Quantitect Reverse Transcription Kit. Two independent microarray experiments were performed; each experiment used RNA from four wildtype and four conditional knockout litter matched mouse calvaria.
Project description:To investigate the role of DDX20 in spermatogenesis, we generated Ddx20 flox/flox Ddx4-Cre mice to make a germ cell-specific Ddx20 knockout, and isolated spermatogonia from four-day-old mouse testes by THY1 magnetic beads. We then performed proteomic analysis using protein lysates obtained from THY1 + spermatogonia.
Project description:Spermiogenesis defines the final phase of male germ cell differentiation. Multiple deubiquitinating enzymes have been linked to spermiogenesis, yet the impacts of deubiquitination on spermiogenesis remain poorly characterized. Here, we investigated the function of UAF1 in mouse spermiogenesis and male fertility. We selectively deleted Uaf1 in premeiotic germ cells using Stra8-Cre knock-in mouse strain (Uaf1 sKO), and found that Uaf1 is essential for spermiogenesis and male fertility. We found that UAF1 interacts and colocalizes with USP1 in testes. Conditional knockout of Uaf1 in testes results in disturbed expression and localization of USP1, suggesting that UAF1 regulates spermiogenesis through the function of the deubiquitinating enzyme USP1. We used tandem mass tag-based proteomics to identify differentially expressed proteins and potential underlying mechanisms, and found that conditional knockout Uaf1 in testes results in reduced levels of proteins essential for spermiogenesis. Thus, the UAF1/USP1 deubiquitinase complex is essential for normal spermiogenesis by regulating the levels of spermiogenesis-related proteins.
Project description:Mechanistic insights into MGAT1 loss during spermatogenesis were investigated in germ cells from 22 day males. Gene expression changes induced by deletion of Mgat 1in spermatogonia were determined using the Affymetrix GeneChip Whole Transcript Plus Reagent Kit. We include the expression data obtained from control and conditional knock out Mgat1 mutant mouse testis germ cells . We identified the genes that were differentially expressed (DEGs) in 22 day Mgat1 mutant versus control germ cells, with a significance level of ANOVA p < 0.05 and an absolute fold-change (linear) less than -2 and greater than 2.0.
Project description:We profiled the gene expression/splicing program of normal and hnRNP U-deficient mouse hearts by RNA-seq. RNA-seq profiles of control and Hnrnpu mutant hearts at postnatal day 14. Hnrnpu mutant hearts were generated by breeding the Hnrnpu conditional knockout mice with Ckmm-Cre transgenic mice.
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).
