Dataset Information

Heterozygous and Homozygous Knock-Out of USP7 mouse models [mouse_RNA-seq]

ABSTRACT: RNAseq analysis of USP7 conditional knock-out (cKO) mice. They were designed to flox exon 6 of USP7 and to allow deletion of exon 6 upon expression of Cre recombinase 27. USP7FL/FL mice were bred with Vav1-Cre mice to obtain USP7FL/wt-Vav1-Cre mice (heterozygote). USP7FL/FL-Vav1-Cre mice (homozygote) were obtained via breeding of heterozygous cKO mice.

ORGANISM(S): Mus musculus

PROVIDER: GSE148521 | GEO | 2021/03/09

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:Oncostatin M (OSM) is a member of the interleukin-6 (IL-6) family of cytokines. OSM has been found to have distinct anti-inflammatory and pro-inflammatory properties in various cellular and disease contexts. OSM signals through a receptor complex that includes OSMRbeta and activates downstream signaling through the JAK/STAT, MAPK/ERK and PI3K/AKT pathways. To investigate the anti- and/or pro-inflammatory roles of OSM signaling in the context of adult hematopoiesis, we utilized the readily available Cre-driven conditional knockout Osmrfl/fl mouse model B6;129-Osmrtm1.1Nat/J. This model contains loxP sites flanking exon 2, which is the first coding exon of the Osmr gene. We crossed Osmrfl/fl mice to an interferon-inducible Mx1-Cre allele that is classically utilized to study adult hematopoiesis given its high efficiency of expression and recombination in adult hematopoietic cells. We developed a PCR assay to evaluate recombination of the Osmrfl allele. In Osmrfl/fl Mx1-Cre mice, we observed complete recombination of the Osmrfl allele and loss of exon 2 in hematopoietic (bone marrow) as well as non-hematopoietic (liver, lung, kidney) tissues. To evaluate Osmr mRNA expression downstream of exon 2, real-time PCR was utilized with TaqMan probes that anneal to an amplicon spanning Osmr exons 17 and 18. Using this assay, reduced Osmr transcript was observed in kidney samples from Osmrfl/fl Mx1-Cre mice compared to Mx1-Cre controls. However, Osmr transcript levels were similar in bone marrow, lung, and liver samples from Osmrfl/fl Mx1-Cre compared to Mx1-Cre control mice, suggesting that transcript is being produced despite loss of exon 2. Western blots revealed that liver cells from Osmrfl/fl Mx1-Cre mice had complete loss of OSMR protein. In contrast, bone marrow, kidney, and lung cells from Osmrfl/fl Mx1-Cre mice had reduced levels of OSMR protein. Together, our data suggest that the B6;129-Osmrtm1.1Nat/J model should be utilized with caution as loss of exon 2 of Osmr variably impacts mRNA and protein expression in a tissue-dependent context.

Dataset Information

Heterozygous and Homozygous Knock-Out of USP7 mouse models [mouse_RNA-seq]

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure