RNA-seq of mouse embryonic fibroblasts(MEFs) with knock-out of SERF2 compared to control MEFs
Ontology highlight
ABSTRACT: Serf2 has been identified as a potential modifier of aggregation in the case of several disease-related aggregation-prone proteins, such as amyloid-beta, alpha-synuclein and huntingtin. However, it's endogenous biological function remains unknown. Therefore, we generated Serf2-/- mice using the cre-lox system to explore the endogenous role of Serf2. As the knockout of Serf2 appeared to cause perinatal lethality, we isolated embryonic fibroblasts at embryonic day 13.5 from Serf2+/+ and Serf2-/- embryo's and generated monoclonal cell lines. We compared these cell lines through RNA-sequencing to investigate the role of Serf2 in embryonic development.
Project description:The objective of this study was to decipher the metabolism expressed by Lactobacillus delbrueckii subsp. delbrueckii CIRM-BIA865 during soy juice fermentation using transcriptomics. The whole genome was sequenced, assembled and annotated. CIRM-BIA865 was then used to ferment soy juice to produce a soy-based yogurt. Samples were analysed in kinetics during fermentation, at pH values of 6.5, 6, 5 and 4.6. RNA from CIRM-BIA865 were extracted and sequenced using paired-end Illumina. Reads were mapped using Bowtie2 on previously obtained genome of CIRM-BIA865. No mismatch were allowed. Reads mapped on CDS were counted using htseqcount.List of differentially expressed (DE) genes between two successive sampling times (determined by pH) were generated using DEseq2 with a modified t-test and a p-value adjusted by Bonferoni inferior to 0.05. Fold changes expressed how many times genes were induced along the fermentations.
Project description:We investigated the genomic binding profiles of the transcription factors Pbx1, Prep1, and Sp2 in mouse embryonic fibroblasts using ChIPseq and ChIPexo. Refer to E-MTAB-2970 and E-MTAB-994 for KO and IgG Controls.
Project description:Mouse Ikbkap gene encodes IKAP/Elp1- one of the core subunits of Elongator- and has been implicated in translational regulation. However, a role for IKAP in genome maintenance remains unclear. In this study, we analyze proteins from control and Ikbkap depletion mouse embryonic fibroblasts (MEF) using MS. Using MS-based proteomics, we show that several proteins involved in cancer and DNA damage response were found to be differentially expressed upon Ikbkap depletion.
Project description:The aim of the study was to decipher metabolisms responsible (i) for the peculiar adaptation of L. plantarum during soy juice fermentation and (ii) for the release of aroma compounds, amino and short-chain fatty acid, and metabolites with health-promoting properties in soy yogurt. The strategy was the sequencing and annotation of a strain (L. plantarum CIRM-BIA777, PRJEB77707) able to degrade galacto- oligosaccharides, the sampling of soy yogurt, RNA-seq to identify differentially expressed genes of L. plantarum and corresponding metabolisms throughout the kinetics of fermentation. Acids and volatile compounds were also quantified.
Project description:Serine/threonine kinase 40 (Stk40) was previously identified as a direct target gene of pluripotency-associated transcription factor Oct4 and its overexpression could facilitate differentiation of mouse embryonic stem cells (mESCs) towards the extraembryonic endoderm. Stk40-/- mice are lethal at the perinatal stage, displaying multiple organ failures. However, the molecular mechanisms underlying the physiological functions of Stk40 remain elusive. Here, we report that Stk40 ablation compromises the mesoderm differentiation from mESCs in vitro and in embryos. Mechanistically, Stk40 interacts with both mammalian constitutive photomorphogenic protein 1 (Cop1) and c-Jun, promoting degradation of c-Jun. Consequently, Stk40 knockout leads to c-Jun protein accumulation, which, in turn, might suppress the Wnt signaling activity and impair the mesoderm differentiation process. Overall, this study reveals that Stk40, together with Cop1, represent a novel axis for modulating c-Jun protein levels within an appropriate range during mesoderm differentiation from mESCs. Our finding provides new insight into the molecular mechanism regulating c-Jun protein stability and may have potential for managing related cellular disorders.
Project description:We have mapped binding sites for the histone demethylase, Jmjd2c/Kdm4c/Gasc1, in mouse embryonic fibroblasts (MEFs) and the impact of Jmjd2c depletion on H3K9me3 and H3K36me3 distributions. ChIP-seq was performed using antibodies recognizing Jmjd2c, H3K9me3 or H3K36me3. Chromatin was obtained from conditional Jmjd2c knockout MEFs cultured in the absence or presence of OHT to induce activation of Cre recombinase and loss of Jmjd2c expression.
Project description:Mouse embryonic stem cells (mESCs), derived from pre-implantation blastocyst cells, can be maintained in vitro in defined N2B27 medium supplemented with two chemical inhibitors for GSK3 and MEK (2i) and the cytokine leukemia inhibitory factor (LIF), which act synergistically to promote self-renewal and pluripotency. Many efforts have been devoted to identify genes that promote exit from the pluripotent state and the transition to a primed state of differentiation. One of the first identified players in this process was the Wnt/b-catenin effector TCF7L1 (previously referred to as TCF3), belonging to the family of four TCF/LEF transcription factors, which acts as pro-differentiation factor by repressing pluripotency genes. Of note, there is little evidence that the genetic abrogation of the mechanisms required for the exit from the pluripotent state is sufficient to enable self-renewal in the absence of 2iL. Here, we found that complete loss-of-function of Tcf7, Lef1, Tcf7l1 and Tcf7l2, the genes encoding for the four TCF/LEF transcription factors, (refered to as qKO) allows mESCs to become fully 2iL-independent and to propagate in basal N2B27. To understand the genetic program that allows qKO cells to achieve 2iL-independent self-renewal, we performed RNA sequencing (RNA-seq) of qKO and wild type mESCs.
Project description:E4F1 is a ubiquitously expressed zinc-finger protein of the Gli-Kruppel family that was first identified, more than 30 years ago, as a cellular target of the adenoviral oncoprotein E1A13S (Ad type V), required for transcriptional regulation of adenoviral genes. In order to decipher E4F1 cellular target genes, we performed chromatin immunoprecipitation of endogenous E4F1 in primary and in p53KO, Ha-RasV12-transformed MEFs. Both input and immunoprecipitated DNA were exhaustively sequenced and mapped on the mouse genome (mm9). Peak detection has been achieved by combining two peak calling algorithms. Intersection of the two E4F1 peak lists on each cell line were considered as E4F1 chromatin bound regions. Genome-wide mapping of E4F1 binding in mouse embryonic fibroblasts.
Project description:Transcription profiling by array of wild type and p300 KIX/KIX; CBP +/KIX primary mouse embryonic fibroblasts (MEFs) transduced with either MSCV-c-Myb_IRES-GFP or MSCV-IRES-GFP retrovirus to determine the effect of mutating the KIX domain of CBP and p300 on c-Myb regulated gene expression. CBP KIX mutation (MGI:3578129) and p300 KIX mutation (MGI:3578128).