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Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Mus musculus

PROVIDER: GSE123422 | GEO | 2019/10/30

REPOSITORIES: GEO

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Tagger-A Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice.

Project description:A deep understanding of how regulation of the multiple levels of gene expression in mammalian tissues give rise to complex phenotypes has been impeded by cellular diversity. A handful of techniques were developed to tag-select nucleic acids of interest in specific cell types, thereby enabling their capture. We expanded this strategy by developing the Tagger knock-in mouse line bearing a quad-cistronic transgene combining enrichment tools for nuclei, nascent RNA, translating mRNA, and mature microRNA (miRNA). We demonstrate that Tagger can capture the desired nucleic acids, enabling multiple omics approaches to be applied to specific cell types in vivo using a single transgenic mouse line.

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Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice [Ribo-Tag-RNA-seq]

Project description:We developed a knock-in mouse line that combines four enrichment tools to study gene regulatory networks of the brain and other complex tissues. As a result, chromatin, nascent RNA, translating mRNA and mature miRNA can be purified from specific cell populations. Specifically, we developed "Tagger" – a Cre and/or Flp recombinase-dependent mouse model that expresses four proteins to enable purification of distinct populations of cellular nucleic acids from specific cell types. Upon activation by cell type-directed recombinases, cells of a desired population express: 1) exogenous uracil phoshoribosyltransferase (UPRT) for tagging nascent total RNA with thiol groups upon administration of 4-thiouracil (4TU, TU-Tag), 2) epitope-tagged large ribosomal subunit protein 22 (Rpl22) to co-immunoprecipitate (co-IP) ribosome-bound mRNA (RiboTag), 3) epitope-tagged Argonaute 2 (Ago2, a component of the RNA-induced silencing complex) to co-IP mature miRNA (AgoTag), and 4) nucleus-directed mKate2 red fluorescent protein, providing good tissue penetrating transmission properties for in vivo imaging and a fluorescent nuclear tag (NucTag) for FACS enrichment of nuclei for chromatin studies.

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Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice [Nuc-Tag-MeDIP-seq]

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Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice [TU-Tag-RNA-seq]

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Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice

Project description:Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice

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US28: HCMV's Swiss Army Knife.

Project description:US28 is one of four G protein coupled receptors (GPCRs) encoded by human cytomegalovirus (HCMV). The US28 protein (pUS28) is a potent signaling molecule that alters a variety of cellular pathways that ultimately alter the host cell environment. This viral GPCR is expressed not only in the context of lytic replication but also during viral latency, highlighting its multifunctional properties. pUS28 is a functional GPCR, and its manipulation of multiple signaling pathways likely impacts HCMV pathogenesis. Herein, we will discuss the impact of pUS28 on both lytic and latent infection, pUS28-mediated signaling and its downstream consequences, and the influence this viral GPCR may have on disease states, including cardiovascular disease and cancer. We will also discuss the potential for and progress towards exploiting pUS28 as a novel therapeutic to combat HCMV.

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Project description:Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice [TU-Tag-RNA-seq]

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Project description:Tagger–a Swiss army knife for multiomics to dissect cell type-specific mechanisms of gene expression in mice [Ribo-Tag-RNA-seq]

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