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Stau2 RIP-Chip in Mouse E13-14 Cortex

ABSTRACT: The double-stranded RNA binding protein Staufen2 (Stau2) is asymmetrically localized and segregated during asymmetric cell divisions in the developing mouse cortex and promotes intermediate progenitor cell fate. We carry out RNA immunoprecipitation with a Stau2 specific antibody, followed by microarray analysis to identify Stau2 assoicated RNAs that may be shuttled asymmetrically into one daughter cell thus affecting cell fate.

ORGANISM(S): Mus musculus

PROVIDER: GSE38222 | GEO | 2012/12/21

SECONDARY ACCESSION(S): PRJNA167456

REPOSITORIES: GEO

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Stau2 RIP-Chip in Mouse E13-14 Cortex

Project description:The double-stranded RNA binding protein Staufen2 (Stau2) is asymmetrically localized and segregated during asymmetric cell divisions in the developing mouse cortex and promotes intermediate progenitor cell fate. We carry out RNA immunoprecipitation with a Stau2 specific antibody, followed by microarray analysis to identify Stau2 assoicated RNAs that may be shuttled asymmetrically into one daughter cell thus affecting cell fate. Cortex was dissected from E13-14 timed-pregnant Swiss Webster Mice (Taconic) and RNA immunoprecipitation followed by microarray analysis was carried out to identy Stau2 associated RNA cargos.

2012-12-21 | E-GEOD-38222 | biostudies-arrayexpress

Altered Glutamate Receptor Ionotropic Delta Subunit 2 expression in Stau2 deficient cerebellar Purkinje cells in the adult brain

Project description:Staufen2 (Stau2) is an RNA-binding protein that is involved in dendritic spine morphogenesis and function. Several studies have recently investigated the role of Stau2 in the regulation of its neuronal target mRNAs, with particular focus on the hippocampus. Here, we provide evidence for Stau2 expression and function in cerebellar Purkinje cells. We show that Stau2 downregulation (Stau2GT) lead to an increase of the glutamate receptor ionotropic delta subunit 2 (GluD2) in Purkinje cells when animals performed physical activity by voluntary wheel running. Furthermore, Stau2GT mice showed lower performance in motor coordination assays but enhanced motor learning abilities, concomitantly with an increase in dendritic GluD2 expression. Together, our results suggest a novel role of Stau2 in Purkinje cell synaptogenesis in the mouse cerebellum

2019-04-23 | GSE126996 | GEO

Chromatin immunoprecipitation profiling of human HEK293 cells transiantly transfected with plasmids coding for Staufen2-HA isoforms

Project description:In human cells, Staufen2 is a double-stranded RNA-binding protein involved in several cellular functions. Although 51% identical to Staufen1, these proteins are nevertheless found in different RNA particles. In addition, differential splicing events generate Staufen2 isoforms that only differ at their N-terminal extremities. We used a genome wide approach to identify and compare the mRNA targets of mammalian Staufen2 isoforms. The mRNA content of Staufen mRNPs was identified by probing DNA microarrays with probes derived from mRNAs isolated from immunopurified Staufen2-containing complexes following transfection of HEK293T cells with Stau2-HA (59kDa) or Stau2-HA (62kDa) expressors. Our results indicate that 11% of the cellular RNAs expressed in HEK293T cells are found in Stau2-containing mRNPs. There is a predominance of mRNAs involved in cell metabolism, transport, transcription, regulation of cell processes and catalytic activity. Experiment Overall Design: HEK293 cells were transiantly transfected with plasmids coding for Staufen2-HA isoforms. Cell extracts were immunoprecipitated using anti-HA antibodies and co-immunoprecipitated mRNAs were purified and used to hybridize microarrays. As control, cell extract from mock transfected cells were used.

2008-06-16 | E-GEOD-8437 | biostudies-arrayexpress

IP Staufen2

2008-01-18 | GSE8437 | GEO

Stau2 knockdown in human bcCML cells (K562)"

Project description:STAU2 knockdown in human K562 cells

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Sequencing of RNA Isolated Through RNA Immunoprecipitation (RIP) of Murine Cortical Tissue Using STAU2 Antibody

Project description:We report here the sequencing and analysis of RNA isolated through RNA immunoprecipitation (RIP) of embryonic mouse cortical tissue. STAU2 antibodies were used to perform these RIPs at four different ages, and the RNA was compared against matching input samples. This allowed characterization of putative STAU2 binding sites in different RNA classes over the course of development, important as STAU2 is a known double-stranded RNA binding protein with a role in asymmetric division of neural progenitor cells.

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Effect of Stau2 knockdown on H3K4 methylation in human bcCML cells (K562)

Project description:Evaluating the effect of Stau2 knockdown on H3K4me1, H3K4me2, H3K4me3 in K562 cells

2020-04-21 | GSE142307 | GEO

Rattus novegicus cerebral cortex hiCLIP mapping of STAU2 binding sites

Project description:Using an improved hiCLIP protocol (Lee & Chakrabarti et al.), we mapped the binding sites of STAU2 in rat cortex and their secondary structure context.

2021-12-04 | E-MTAB-11201 | biostudies-arrayexpress

iCLIP analysis of RNA binding sites of Stau2 protein in mouse brain cells

Project description:Stau2 iCLIP of mouse brain was performed to identify RNA binding sites of Stau2 protein in mouse brain cells. The experiment was performed in triplicate, and each of the replicates was split into two separate halves at the cDNA stage, which together led to 6 separate datasets. The iCLIP protocol includes the following steps; Embryonic day 18 whole mouse brain was dissociated and irradiated with UV-C light, and the cells were then lysed using a buffer containing detergents. The RNAs were partially digested, and Stau2 and the cross-linked RNA fragments were immunoprecipitated using anti-Stau2 antibody. A DNA adaptor was then ligated to the RNA fragments and the cross-linked RNAs were further purified by SDS-PAGE and nitrocellulose membrane transfer. The RNAs were extracted from the membrane by proteinase K treatment, and converted into a high-throughput DNA sequencing compatible library by reverse transcription and PCR. For further details, see the methods of the associated manuscript. Note that the sequence reads start with (3 nucleotides of unique molecular identifiers) + (4 nucleotides of experimental barcode) + (2 nucleotides of unique molecular identifiers) followed by the sequence of the cross-linked RNA fragments.

2017-08-02 | E-MTAB-5703 | biostudies-arrayexpress

An In Vivo Genome-Wide CRISPR Screen Identifies the RNA-Binding Protein Staufen2 as a Key Regulator of Myeloid Leukemia

Project description:While Chronic Myelogenous Leukemia (CML) is generally well controlled with Imatinib and other kinase inhibitors, blast crisis CML (bcCML) continues to be resistant to current therapies and remains highly lethal. In this study, we identify the double stranded RNA binding protein Stau2 as a as key new dependency of bcCML. To define the impact of Staufen2 in myeloid leukemia, we developed a germline knockout with targeted loss of its RNA binding domains. Genetic deletion of Staufen2 led to a profound decrease in bcCML growth and propagation, and markedly improved overall survival in mouse models. Further, Staufen2 was also expressed and required for growth of primary bcCML as well as de novo AML patient samples, indicating a conserved utilization across human disease. Finally, an integrated computational analysis of eCLIP, RNA-Seq analysis and the functional genomic CRISPR screen identified Staufen2 as a regulator of a remarkable network of broad acting chromatin modifiers, resulting in changes in global histone methylation patterns.

2020-04-23 | GSE134971 | GEO

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