{"database":"GPMDB","file_versions":[],"scores":{"citationCount":0,"reanalysisCount":0,"viewCount":33,"searchCount":5},"additional":{"omics_type":["Other"],"submitter":["Hagen L, et al."],"instrument_platform":["Instrument"],"disease":["Not Available"],"brenda_tissue":["Not available"],"species":["Homo_sapiens_viruses, Human_female"],"submitter_mail":["geir.slupphaug@ntnu.no"],"publication":["25448019"],"submitter_affiliation":["Department of Cancer Research and Molecular Medicine and PROMEC Core Facility for Proteomics and Metabolomics, Norwegian University of Science and Technology"],"model":["http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005249","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005266","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005264","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005265","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005265","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005264","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005262","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005263","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005269","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005266","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005268","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005267","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005250","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005259","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005253","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005254","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005251","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005252","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005257","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005258","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005255","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005256","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005260","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310005261","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005261","http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32320005260"],"cell_type":["Not available"],"repository":["GPMDB"],"pubmed_abstract":["Transient transfection of mammalian cells with plasmid expression vectors and chemical transfection reagents is widely used to study protein transport and dynamics as well as phenotypic alterations mediated by the overexpressed protein. Despite the undisputed impact of this technique, surprisingly little is known about the cellular effects mediated by the transfection process per se. Conceivably, off-target effects could have implications upon proteins or processes being studied and understanding the molecular pathways affected would add value to the interpretation of experimental observations subsequent to cell transfection. Here we have used a SILAC-based proteomic approach to study differentially expressed proteins after transfection of HeLa cells with ECFP vector using a commonly employed non-liposome based transfection reagent, Fugene®HD. Whereas the transfection reagent itself mediated minimal effects upon protein expression, 11 proteins were found to be significantly upregulated after transfection, all of which were associated with an interferon type I/II response. The upregulated proteins might potentially inflict major cellular processes such as RNA splicing, chromatin remodeling, post-translational protein modification and cell cycle control. The results were validated by western analysis as well as quantitative RT-PCR and this demonstrated that an essentially identical response was induced in HeLa by transfection using an empty pUC18 vector, which does not contain a mammalian virus promoter, as well as a liposome-based transfection reagent, Lipofectamine(TM)2000. Notably, no induction of the interferon response was observed in HEK293 cells, suggesting that these cells might be preferable to HeLa to avoid undesired off-target effects in transfection studies encompassing interferon-signaling and antiviral responses."],"pubmed_title":["Off-target responses in the HeLa proteome subsequent to transient plasmid-mediated transfection."],"pubmed_authors":["Hagen Lars L,Sharma Animesh A,Aas Per Arne PA,Slupphaug Geir G,","Hagen Lars L, Sharma Animesh A, Aas Per Arne PA, Slupphaug Geir G"],"name_synonyms":["HeLa, RCB0007, transfect, Transfections., Proteomes, transient structure"],"description_synonyms":["projections, data, RNA, MPFD, posttranslational modification, anatomical protrusion, chromatin remodelling, CFTD, FuGene 6, lamellae, anatomical process, AUTSX5, Proteins, ACTA, lamina, DXS648, flanges, Gene, regulation of cell cycle transition, CCN3, QM, process of organ, Cell, Splicings, protrusion, polypeptide, lamella, RNA Splicings, Splicing, 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293T Cell, LF 2000, 293 Cells, biological signaling, Transmembrane, Trafficking, posttranslational modification, chromatin remodelling, AU023367, enzyme transport, experimental, determination, lamellae, number, Human Embryonic Kidney Cell Line 293, Gene, regulation of cell cycle transition, RT-PCR, HEK293 Cell, Protein Transports, process of organ, Xt, presence, 293T, protrusion, lamella, Readability, Messenger, responsivity, chromatin modeling, Gene Products, Virus, HEK 293, Animal, Type I, ADHD, chromatin modelling, study, reactivity, Targeting, Animal Virus, Interferons, posttranslational amino acid modification, Protein Trafficking, Gated Protein Transport, methods, Viruses, cell, experimental section, Interferons Type I, Protein Translocation, HeLa, proteins, ridges, posttranslational protein modification, Protein Sortings, Vira, papilla, signaling process, transient structure, Protein Transport, Transmembrane Protein Transport, LF-2000, 293T Cells, RCB0007, associated, laminae, 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modification, Type I Interferons, Cells, HEK, processus, Animal Viruses, Attention Deficit Hyperactivity Disorder, assay, response, quantitative, reverse transcription polymerase chain reaction, E430016J11Rik, Interferon, Transmembrane Protein Transports, RWDD5, presence or absence in 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File: 140508_SILAC_transfect_HeavyReagent_LightNormal_2.mzml. Published as part of Biochim Biophys Acta. 2014 Nov 5;1854(1):84-9  . From the Abstract: {{i}} Here we have used a SILAC-based proteomic approach to study differentially expressed proteins after transfection of HeLa cells with ECFP vector using a commonly employed non-liposome based transfection reagent, Fugene HD. Whereas the transfection reagent itself mediated minimal effects upon protein expression, 11 proteins were found to be significantly upregulated after transfection, all of which were associated with an interferon type I/II response. The upregulated proteins might potentially inflict major cellular processes such as RNA splicing, chromatin remodeling, post-translational protein modification and cell cycle control. {{/i}}","dates":{"submission":"2014-12-12"},"accession":"GPM32320005268","cross_references":{"pubmed":["25448019"],"Pride":["PXD001278"],"pride":[],"Pride Archive":["PXD001278"]}}