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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Temporal Endogenous Gene Expression Profiles in Response to Polymer-Mediated Transfection and Profile Comparison to Lipid-Mediated Transfection

ABSTRACT: The current understanding of the molecular factors underlying polyethylenimine(PEI)-mediated transfection is largely unknown. Cationic PEI was used to transfer GFP transgene to HEK293T cells. FACS separation of transfected (GFP positive), untransfected (GFP negative), and untreated cells enabled gene expression profiles to be obtained using Affymetrix® HG-U133A 2.0 microarrays for each cell population. Gene profiles were differentially compared for each population combination. Control (untreated), transfected (GFP positive), and untransfected (GFP negative) RNA samples were purified from HEK293T cells after FACS isolation. Treated experiments were repeated on triplicate days to run 3 microarrays for each cell population. Controls were run on duplicate to give 2 microarrays for each population.

ORGANISM(S): Homo sapiens

SUBMITTER: Timothy Martin

PROVIDER: E-GEOD-59665 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Temporal Endogenous Gene Expression Profiles in Response to Lipid-Mediated Transfection

Project description:The current understanding of the molecular factors underlying LF2000-mediated transfection is largely unknown. Cationic LF2000 gene delivery system was used to transfer GFP transgene to HEK293T cells. FACS separation of transfected (GFP positive), untransfected (GFP negative), and untreated cells enabled gene expression profiles to be obtained using Affymetrix® HG-U133A 2.0 microarrays for each cell population. Gene profiles were differentially compared for each population combination. Control (untreated), transfected (GFP positive), and untransfected (GFP negative) RNA samples were purified from HEK293T cells after FACS isolation. Treated experiments were repeated on triplicate days to run 3 microarrays for each cell population. Controls were run on duplicate to give 2 microarrays for each population.

2014-07-23 | E-GEOD-59666 | biostudies-arrayexpress

Network Analysis of HEK293T Gene Expression Profile Modulation During Nonviral Gene Delivery

Project description:The current understanding of the molecular factors underlying PEI-mediated transfection is largely unknown. The cationic polyethyleneimine (PEI) polymer gene delivery system was used to transfer the GFP gene to HEK293T cells. FACS separation of transfected (GFP positive), untransfected (GFP negative), and untreated cells enabled gene expression profiles to be obtained using Affymetrix® HG-U133A 2.0 microarrays for each cell population. Gene profiles were differentially compared for each population combination. Control (untreated), transfected (GFP positive), and untransfected (GFP negative) RNA samples were purified from HEK293T cells after FACS isolation. The experiment was repeated on triplicate days to run 3 microarrays for each cell population. Only 2 untreated replicates are included in this submission.

2012-06-02 | E-GEOD-38422 | biostudies-arrayexpress

Temporal Endogenous Gene Expression Profiles in Response to Polymer-Mediated Transfection and Profile Comparison to Lipid-Mediated Transfection

Project description:The current understanding of the molecular factors underlying polyethylenimine(PEI)-mediated transfection is largely unknown. Cationic PEI was used to transfer GFP transgene to HEK293T cells. FACS separation of transfected (GFP positive), untransfected (GFP negative), and untreated cells enabled gene expression profiles to be obtained using Affymetrix® HG-U133A 2.0 microarrays for each cell population. Gene profiles were differentially compared for each population combination.

2014-07-23 | GSE59665 | GEO

Network Analysis of HEK293T Gene Expression Profile Modulation During Nonviral Gene Delivery

2012-06-03 | GSE38422 | GEO

Osmotic stress induces phosphorylation of histone H3 at threonine 3 in pericentromeric regions of Arabidopsis thaliana [expression]

Project description:Histone phosphorylation plays key roles in stress-induced transcriptional reprogramming in metazoans but its function(s) in land plants has remained relatively unexplored. Here we report that an Arabidopsis mutant defective in At3g03940 and At5g18190, encoding closely related Ser/Thr protein kinases, shows pleiotropic phenotypes including dwarfism and hypersensitivity to osmotic/salt stress. The double mutant has reduced global levels of phosphorylated histone H3 threonine 3 (H3T3ph), which are not enhanced, unlike the response in the wild type, by drought-like treatments. Genome-wide analyses revealed increased H3T3ph, slight enhancement in trimethylated histone H3 lysine 4 (H3K4me3), and a modest decrease in histone H3 occupancy in pericentromeric/knob regions of wild type plants under osmotic stress. However, despite these changes in heterochromatin, transposons and repeats remained largely transcriptionally repressed. In contrast, this reorganization of heterochromatin was mostly absent in the double mutant, which even under normal conditions exhibited lower H3T3ph levels in pericentromeric regions, and a few transposons and repeat sequences showed modest transcriptional activation. Interestingly, within actively transcribed protein-coding genes, H3T3ph density was minimal in 5’ genic regions, coincidental with a peak of H3K4me3 accumulation. This pattern was not affected in the double mutant, implying the existence of additional H3T3 protein kinases in Arabidopsis. Our results suggest that At3g03940 and At5g18190 are involved in the phosphorylation of H3T3 in pericentromeric/knob regions and that this repressive epigenetic mark may be important for maintaining proper heterochromatic organization and, possibly, chromosome function(s). Columbia-0 and double mutant at3g03940/at518190 knockdown plants were grown in 12 hr light for 3 weeks in pots in well-watered state. RNA was isolated from rosettes in triplicate for analysis on microarray.

2015-07-22 | E-GEOD-68439 | biostudies-arrayexpress

Effect of Protease-resistant PML-RARα on the leukemogenic potential in a mouse model of Acute Promyelocytic Leukemia

Project description:Previous studies in our laboratory demonstrated that the azurophil granule protease neutrophil elastase (NE) cleaves PML-RARA (PR), the fusion protein that initiates acute promyelocytic leukemia (APL). Further, NE deficiency reduces the penetrance of APL in a murine model of this disease. We therefore predicted that NE-mediated PR cleavage might be important for its ability to initiate APL. To test this hypothesis, we generated a mouse expressing NE-resistant PR. These mice developed APL indistinguishable from wild type PR, but with significantly reduced latency (median leukemia-free survival of 274 days versus 473 days for wild type PR, p<0.001). Resistance to proteolysis may increase the abundance of full length PR protein in early myeloid cells, and our previous data suggested that non-cleaved PR may be less toxic to early myeloid cells. Together, these effects appear to increase the leukemogenicity of NE-resistant PR, contrary to our previous prediction. We conclude that NE deficiency may reduce APL penetrance via indirect mechanisms that are still NE dependent. Keywords: Time course U937 cells were transfected with one of three tagged constructs GFP,GFP-PR,GFP-PR-2VR sorted for GFP positivity at the indicated time points and harvested for RNA.

2010-09-01 | E-GEOD-21550 | biostudies-arrayexpress

Functionalized carbon nanotube toxicity in Arabidopsis leaves

Project description:Polyethilenimine (PEI) functionalized single walled carbon nanotubes (SWNTs) and carbon-based nanomaterials enable delivery of DNA and RNA in plants. Given the broad-scale use of PEI-functionalized nanomaterials in plants, we sought to investigate the reaction of plant tissues to treatment with PEI-SWNTs and pristine SWNTs. To this end, we infiltrated Arabidopsis thaliana leaves with pristine single walled carbon nanotubes used in RNA silencing applications (SWNTs) and polyethyleneimine-functionalized SWNTs used for plasmid DNA delivery (PEI-SWNTs). We used Arabidopsis as it is a well characterized model plant, for which genomic and detailed gene function information is readily available. To minimize the effects caused by the introduction of exogenous nucleic acids, in SWNT preparations we used single stranded RNA targeting Green Fluorescent Protein (GFP) with no target sequence in the Arabidopsis genome, and a plasmid that expresses GFP in PEI-SWNT preparations. For our experiments herein, we used ~25-50 fold higher concentrations of SWNTs and PEI-SWNTs compared to standard concentrations used in biomolecule delivery assays. Water-infiltrated plant leaves served as a negative control to distinguish between the SWNT-specific response and the response to the infiltration process itself. We performed RNA sequencing (RNA-seq) with RNA extracted from leaves two days after infiltration to identify changes in the leaf transcriptomic profile in response to the three treatments, compared to non-infiltrated leaves.

2021-12-29 | GSE172278 | GEO

Intracellular Signaling Pathways in Nonviral Gene Delivery: Gene Expression Profiles in Transfected Cells

Project description:Analysis of gene expression changes due to nonviral gene delivery of DNA lipoplexes versus control in human HEK293T cells. Human HEK293T RNA was isolated from control, non-transfected cells (CTR) and transfected (GFP) samples for analysis on microarrays with three biological replicates.

2011-10-04 | E-GEOD-20615 | biostudies-arrayexpress

Gene expression analysis of ectopic cyclin D1-expressing myeloma cells compared to their non expressing counterparts

Project description:We analyzed gene expression profiles of myeloma cells belonging to the group of bas prognosis RPMI 8226 and LP1 expressing either the GFP protein or a cyclin D1-GFP fusion protein Two cell lines RPMI 8226 and LP1. Two clones from each cell line GFP and cyclin D1-GFP. Four biological replicates independently grown and harvested. One replicate per array.

2014-07-23 | E-GEOD-59673 | biostudies-arrayexpress

Pax3 and Pax7 play essential safeguard functions against environmental stress-induced birth defects

Project description:Exposure to environmental teratogenic pollutant leads to severe birth defects. However, the biological events underlying these developmental abnormalities remain undefined. Here we report a molecular link between an environmental stress response pathway and key developmental genes during craniofacial development. In our study, we focused on the development of the facial prominences at E11.5. To do so, we compared the transcriptomes of mutant embryos (*Pax3Pax3-ERD/GFP *called DM in the microarray samples) to the one of control embryos (*Pax3GFP/+ *called GFP in the sample names). These are knock-in genetic models described in Bajard et al., 2006 and Relaix et al., 2005. In both of them a cassette coding for the GFP is replacing one allele of the Pax3 gene. The Pax3-ERD allele is a conditional one that drives the expression of the dominant negative form of Pax3 (Pax3-ERD) composed of the Pax3 DNA binding domain fused to the engrailed repressor domain (ERD) upon activation of a Cre recombinase. In this study, the Cre was driven by the zygote specific PGK enhancer. Strikingly, mutant mice with impaired Pax3/7 function display severe craniofacial defects. We show these are associated with an up-regulation of the signaling pathway mediated by the Aryl hydrocarbon Receptor (AhR), the receptor to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), revealing a genetic interaction between Pax3 and AhR signaling. Activation of AhR signaling in Pax3-deficient embryos drives facial mesenchymal cells out of the cell cycle through the up-regulation of p21 expression. Accordingly, inhibiting AhR activity rescues the cycling status of these cells and the facial closure of Pax3/7 mutants. Together, our findings demonstrate that the regulation of AhR signaling by Pax3/7 is required to protect against TCDD/AhR-mediated teratogenesis during craniofacial development.

2015-04-06 | E-MTAB-3238 | biostudies-arrayexpress

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