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Transcriptome profiling of HEK 293T cells depleted of endogenous miRNAs

ABSTRACT: To assess the impact of AdV-VP55 mediated degredation of host miRNAs on the cellular transcriptome.

ORGANISM(S): Homo sapiens

PROVIDER: GSE73718 | GEO | 2016/01/30

SECONDARY ACCESSION(S): PRJNA297763

REPOSITORIES: GEO

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Small RNA deep sequencing of HEK 293T cells treated with an Adeno vector encoding the Vaccinia virus VP55 protein

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Small RNA deep sequencing of HEK 293T cells treated with an Adeno vector encoding the Vaccinia virus VP55 protein

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2016-08-03 | E-GEOD-74088 | biostudies-arrayexpress

Transcriptome profiling of murine lungs after treatment with vector to deplete endogenous miRNAs

Project description:To assess the impact of AdV-VP55 mediated degredation of host miRNAs on the murine whole lung transcriptome profile Non-replicateive Adeno vectors were delivered intranasally to 6-8 week old C57BL/6 mice for 48hrs. Whole lung tissue was isolated and used for transcriptome profiling.

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Transcriptome profiling of HEK 293T Dicer -/- cells with or without VP55

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Transcriptome profiling of HEK 293T cells and BJ foreskin fibroblasts depleted of endogenous miRNAs

Project description:This SuperSeries is composed of the SubSeries listed below.

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Transcriptome profiling of HEK 293T cells and BJ foreskin fibroblasts depleted of endogenous miRNAs

Project description:Transcriptome profiling of HEK 293T cells and BJ foreskin fibroblasts depleted of endogenous miRNAs

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Tumorigenic effects of TLX overexpression in HEK 293T cells.

Project description:BackgroundThe human orphan receptor TLX (NR2E1) is a key regulator of neurogenesis, adult stem cell maintenance, and tumorigenesis. However, little is known about the genetic and transcriptomic events that occur following TLX overexpression in human cell lines.AimsHere, we used cytogenetics and RNA sequencing to investigate the effect of TLX overexpression with an inducible vector system in the HEK 293T cell line.Methods and resultsConventional spectral karyotyping was used to identify chromosomal abnormalities, followed by fluorescence in situ hybridization (FISH) analysis on chromosome spreads to assess TLX DNA copy number. Illumina paired-end whole transcriptome sequencing was then performed to characterize recurrent genetic variants (single nucleotide polymorphisms (SNPs) and indels), expressed gene fusions, and gene expression profiles. Lastly, flow cytometry was used to analyze cell cycle distribution. Intriguingly, we show that upon transfection with a vector containing the human TLX gene (eGFP-hTLX), an isochromosome forms on the long arm of chromosome 6, thereby resulting in DNA gain of the TLX locus (6q21) and upregulation of TLX. Induction of the eGFP-hTLX vector further increased TLX expression levels, leading to G0-G1 cell cycle arrest, genetic aberrations, modulation of gene expression patterns, and crosstalk with other nuclear receptors (AR, ESR1, ESR2, NR1H4, and NR3C2). We identified a 49-gene signature associated with central nervous system (CNS) development and carcinogenesis, in addition to potentially cancer-driving gene fusions (LARP1-CNOT8 and NSL1-ZDBF2) and deleterious genetic variants (frameshift insertions in the CTSH, DBF4, POSTN, and WDR78 genes).ConclusionTaken together, these findings illustrate that TLX may play a pivotal role in tumorigenesis via genomic instability and perturbation of cancer-related processes.

| S-EPMC7941458 | biostudies-literature

small RNA-seq in HEK-293t cells

Project description:The effect of NF90 on the abundance of miRNAs was measured using small RNA-seq.

2020-05-01 | GSE132335 | GEO

Cloning, Expression and Characterization of Zebra Fish Ferroportin in Hek 293T Cell Line.

Project description:BACKGROUND: Ferroportin (Fpn), a regulator of iron homeostasis is a conserved membrane protein that exports iron across the enterocytes, macrophages and hepatocytes into the blood circulation. Fpn has also critical influence on survival of microorganisms whose growth is dependent upon iron, thus preparation of Fpn is needed to study the role of iron in immunity and pathogenesis of micoorganisms. METHODS: To prepare and characterize a recombinant ferroportin, total RNA was extracted from Indian zebrafish duodenum, and used to synthesize cDNA by RT-PCR. PCR product was first cloned in Topo TA vector and then subcloned into the GFP expression vector pEGFP-N1. The final resulted plasmid (pEGFP-ZFpn) was used for expression of Fpn-EGFP protein in Hek 293T cells. RESULTS: The expression was confirmed by appearance of fluorescence in Hek 293 T cells. Recombinant Fpn was further characterized by submission of its predicted amino acid sequences to the TMHMM V2.0 prediction server (hidden Markov model), NetOGlyc 3.1 and NetNGlyc 3.1 servers. The obtained Fpn from indian zebrafish also contained eight transmembrane domains with N- and C-termini inside the cytoplasm and harboured 78 O-glycosylated amino acids. CONCLUSION: The recombinant Fpn from Indian zebra fish was successfully expressed in Hek 293 cell line. Although the discrepancy in two amino acids was observed in our produced Fpn and resulted in an additional O-glycosylation site, but had no effect on the topology of the protein compared to other Fpn described by other researchers. Therefore this construct can be used in future iron studies.

| S-EPMC3481666 | biostudies-literature

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