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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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE59665 | GEO | 2014/07/23

SECONDARY ACCESSION(S): PRJNA255912

REPOSITORIES: GEO

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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. 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-59665 | biostudies-arrayexpress

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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 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

Project description:This study investigated the pulmonary toxicity of Copper oxide nanoparticles (CuO NPs) surface modified with polyethylenimine (PEI) or ascorbate (ASC), was investigated. Rats were exposed nose-only to a fixed exposure concentration of ASC or PEI coated CuO NPs for 5 consecutive days. On day 6 and day 27 post-exposure, pulmonary toxicity markers in bronchoalveolar lavage fluid (BALF) were analyzed and histopathological evaluation of the lungs was performed, along with microarray analyses on whole lung tissue samples.

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Comparative_Interaction_Proteomics_GFP-tagged_MAGEC2_HEK293T

Project description:293T (Human embryonic kidney cells-ATCC CRL-3216) cells were plated at 500,000 cells per 10 cm dish and grown for 48 hours. Cells were then transfected with GFP-MAGEC1 WT or GFP-MAGEC1 L318D in a 3:1 ratio of PEI (polyethylenimine) to DNA. Cells were harvested after 48 hours and lysed in 50 mM Tris pH 7.5, 150 mM NaCl, 1% Triton X-100, protease inhibitor (cOmpleteâ„¢, Mini, EDTA-free (Roche)). Cells were incubated for 25 minutes on ice and then sonicated for 5 minutes at 4Â°C. Cells were centrifuged at 14,000 rpm for 20 minutes at 4 Â°C. 500 ug lysate was then added to GFP-TrapÂ® Agarose resin (ChromoTek) and samples were incubated rotating at 4Â°C for 2 hours. Following washing of the beads with 50 mM Tris pH 7.5, 150 mM NaCl, proteins were eluted by the addition of 2x Laemmli buffer and boiled for 10 minutes at 95 Â°C. Samples were spun down and the supernatants were sent for mass spectrometry analysis as described below. Experiments were conducted in triplicate.

2024-02-20 | PXD047849 | Pride

Gene expression after transfection of IGF-II specific siRNAs

Project description:Total RNA was isolated from HuH-7 cells after transfection of IGF-II specific siRNAs. Gene expression profiling was performed using the Affymetrix Human Genome U133A 2.0 Arrays. The raw data were analysed using mixed model ANOVA. Keywords: transfection with siRNA Cells transfected with two independent siRNAs (ds1 and ds2) as well as untransfected cells were examined. For each condition biological duplicates were used for gene expression profiling

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Transfection with Glucuronidase A affects general proteome pattern in Giardia lamblia trophozoites

Project description:Giardia lamblia is an important causative agent of persistent diarrhea in humans, domestic animals, and cattle. Basic research is usually performed with the strain WBC6 and includes genetic manipulations such as transfections. Here, we investigate how transfection with a plasmid causing stable expression of a foreign gene affects the whole proteome pattern. Using shotgun mass spectrometry, we compare the proteomes of untransfected trophozoites to trophozoites transfected with Escherichia coli glucuronidase A (GusA). Besides GusA, which is detected in the transfected trophozoites only, the proteomes of untransfected and transfected trophozoites differ by 132 differentially expressed proteins. In particular, transfection induces antigenic variation. Since transfection causing stable expression affects the proteome pattern, transfection experiments should take into account this effect. Due to a unique peptide panel, GusA is an example for a suitable internal standard for experiments involving transfected cells.

2021-01-07 | PXD022565 | Pride

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