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Total Gene expression analysis of H3f3b conditional knockout MEFs

ABSTRACT: Total gene expression analysis was performed on CRE induced conditional knockout E12.5 MEFs relative to GFP infected control MEFs. Intent was to analyze the role of H3f3b in overall gene expression.

ORGANISM(S): Mus musculus

PROVIDER: GSE35301 | GEO | 2013/04/11

SECONDARY ACCESSION(S): PRJNA156025

REPOSITORIES: GEO

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Total Gene expression analysis of H3f3b conditional knockout MEFs

Project description:Total gene expression analysis was performed on CRE induced conditional knockout E12.5 MEFs relative to GFP infected control MEFs. Intent was to analyze the role of H3f3b in overall gene expression. For conditional KO, three lines of H3f3b FL/FL E12.5 MEFs (cKO1, cKO2 and pcKO1) were transduced with Cre retroviruses and compared to their respective lines of MEFs transduced solely with GFP vector (as control). Total RNA was isolated for gene analysis.

2013-04-11 | E-GEOD-35301 | biostudies-arrayexpress

Total Gene expression analysis of H3f3b null and WT MEFs

2013-04-10 | GSE35302 | GEO

Total Gene expression analysis of H3f3b null and WT MEFs

Project description:Total gene expression analysis was performed on constitutive H3f3b KO E12.5 MEFs relative to WT littermates. Intent was to analyze the role of H3f3b in overall gene expression. Constitutive KO mice were obtained by mating H3f3b Fl/Fl mice to Zp3Cre mice. Heterozygous offspring were time mated and embryos were isolated 12.5 days after. MEFs were cultured transduced with a stable pBabe puro vector and total RNA was isolated from H3f3b KO and WT MEFs.

2013-04-10 | E-GEOD-35302 | biostudies-arrayexpress

Total Gene expression analysis of H3f3b constitutive knockout testis RNA

Project description:Total gene expression analysis was performed on RNA from testes extracted from two litters of constitutive homozygous and heterozygous H3f3b knockout mice compared to WT littermates. Constitutive knockout mice were obtained by mating H3f3b Fl/Fl mice to Zp3Cre mice. Heterozygous knockout mice were crossed until homozygous knockout mice were obtained. Testes were surgically removed from 8 week old homozygous knockouts, heterozygous knockouts and WT, and RNA was extracted from one testis from each mouse for total gene expression.

2014-07-31 | E-GEOD-35303 | biostudies-arrayexpress

Total Gene expression analysis of H3f3b constitutive knockout testis RNA

2014-07-31 | GSE35303 | GEO

Effect of depletion of h3f3b on gene expression in vascular smooth muscle cells (VSMCs).

Project description:To investigate the function H3f3b in the regulation of VSMC phenotypic transition, we infect VSMCs with shNT or shH3f3b virus.

2024-04-06 | GSE229125 | GEO

RNA sequencing analysis of Nras mutant MEFs

Project description:Purpose: Emerging evidence in the field of RAS biology is beginning to illustrate that RAS mutations are functionally distinct. To elucidate functional differences between NRAS mutants we performed RNA-sequencing on mouse embryonic fibroblasts (MEFs) expressing different NRAS codon 61 variants from the endogenous gene locus Methods: MEFs were isolated from Tyr::CRE-ERT2 LSL-Nras mouse models containing different Nras variants in the endogeous gene locus. NRAS expression was induced in MEFs using an Adenoviral Cre delivery system. RNA was isolated from MEFs six days post-NRAS induction and submitted for RNA-sequencing Results: Differential expression and gene set enrichment analysis (GSEA) between NRAS variants revealed enrichment of transcripts associated with cellular proliferation between all mutant Nras alleles versus wild-type Nras. Further, MEFs expressing NRAS mutants capable of driving melanoma formation were enriched for transcripts associated with RAS-MYC signaling axis. Conclusion: While all NRAS mutants are capable of enhancing cellular proliferation versus wild-type NRAS, the melanomagenic potential of these mutants correlates with the ability to enhance activation of the RAS-MYC signaling axis

2022-04-30 | GSE162124 | GEO

A knockout-first model of H3f3a gene targeting leads to developmental lethality

Project description:Histone variant H3.3 is encoded by two genes, H3f3a and H3f3b, which can be expressed differentially depending on tissue type. Previous work in our lab has shown that knockout of H3f3b causes some neonatal lethality and infertility in mice, and chromosomal defects in mouse embryonic fibroblasts (MEFs). Studies of H3f3a and H3f3b null mice by others have produced generally similar phenotypes to what we found in our H3f3b nulls, but the relative impacts of loss of either H3f3a or H3f3b have varied depending on the approach and genetic background. Here we used a knockout-first approach to target the H3f3a gene for inactivation in C57BL6/J mice. Homozygous H3f3a targeting produced a lethal phenotype at or before birth. E13.5 embryos had some potential morphological differences from WT littermates including smaller size and reduced head size, while an E18.5 null embryo was smaller than its control littermates with several potential defects including small head and brain size as well as small lungs, which would be consistent with a late gestation lethal phenotype. Despite a reduction in H3.3 and total H3 protein levels, the only histone H3 post-translational modification in the small panel assessed that was significantly altered was the unique H3.3 mark phospho-Serine31, which was consistently increased in null neurospheres. H3f3a null neurospheres also exhibited consistent gene expression changes including in protocadherins. Overall, our findings are consistent with the model that there are differential, cell-type-specific contributions of H3f3a and H3f3b to H3.3 functions in epigenetic and developmental processes.

2023-01-03 | GSE216047 | GEO

Proteomic profiling of eIF3a conditional knockout mice

Project description:In this study, a Cre-loxP system was used to generate eIF3a conditional knockout mice. Tandem mass tag (TMT) labeling with LC-MS/MS analysis was used to identify differentially expressed proteins (DEPs) in fat, lungs, skin, and spleen tissue of the eIF3a knockout mice and controls. Bioinformatics analysis was then used to explore the functions and molecular signaling pathways of these protein landscapes.

2023-07-20 | PXD041416 | Pride

Primary murine embryonic fibroblasts (MEFs): Cdk7 wild type versus Cdk7 KO

Project description:Transcriptional profiling of primary Cdk7lox/lox MEFs comparing Adeno-Cre vs Adeno-GFP infected controls. Samples were obtained 7 days after the Adeno-Cre mediated elimination of Cdk7. Goal was to determine the effects of Cdk7 elimination on RNA polII-mediated transcription. Two-condition experiment, Adeno-Cre vs. Adeno-GFP infected Cdk7lox/lox MEFs. Biological replicates: 10 independent primary MEF preparations.

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