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TransfectionPromoter Regions, GeneticBase SequenceMolecular Sequence DataCell LineTranscription, GeneticTranscription FactorsDNA-Binding ProteinsGene Expression RegulationGenes, ReporterRNA, MessengerTumor Cells, CulturedLuciferasesTranscriptional ActivationPlasmidsChloramphenicol O-AcetyltransferaseCells, CulturedDNASp1 Transcription FactorBinding SitesRegulatory Sequences, Nucleic AcidCloning, MolecularNuclear ProteinsEnhancer Elements, GeneticRecombinant Fusion ProteinsHeLa CellsResponse ElementsIschemic Attack, TransientCOS CellsGene ExpressionAmino Acid SequenceGenetic VectorsGene Expression Regulation, EnzymologicProtein BindingTrans-ActivatorsDNA PrimersDNA, ComplementaryCell Line, TumorRecombinant ProteinsSignal TransductionSequence DeletionMutationCell NucleusMutagenesis, Site-DirectedBlotting, Western3T3 CellsReverse Transcriptase Polymerase Chain Reaction5' Flanking RegionDNA FootprintingGreen Fluorescent ProteinsRNA, Small InterferingTime FactorsDown-RegulationCricetinaeBlotting, NorthernNF-kappa BRepressor ProteinsPolyethyleneimineElectroporationGene Transfer TechniquesCercopithecus aethiopsConsensus SequenceProto-Oncogene ProteinsElectrophoretic Mobility Shift AssayPhosphorylationFibroblastsSequence Homology, Nucleic AcidGene Expression Regulation, ViralRestriction MappingDeoxyribonuclease IApoptosisCalciumOligodeoxyribonucleotidesGene Expression Regulation, NeoplasticIntronsReceptors, Cytoplasmic and NuclearEnzyme InhibitorsSp3 Transcription FactorCHO CellsUp-RegulationCell Line, TransformedTranscription Factor AP-1ExonsPolymerase Chain ReactionCarrier ProteinsLuminescent ProteinsKineticsTATA BoxCyclic AMP Response Element-Binding ProteinCell DifferentiationCell DivisionEnzyme ActivationCCAAT-Enhancer-Binding ProteinsGenetic TherapyImmediate-Early ProteinsRats, Sprague-DawleyDose-Response Relationship, DrugProto-Oncogene Proteins c-junReceptors, Retinoic AcidSequence Homology, Amino Acid
ReagentPolyethylenimineRecombinantStableCellsElectroporationProtocolsEfficiencyReagentsProteinsVectorsInvitrogenExpressionFunctionalSufficientMethodsCreationInfluenceMammalianPlasmidStableAssaysProtein expressionAssaySerumCommercially availableYieldsInvestigateClinicalGenesAnalysisAmountsTimeCultureInterestManufacturers
Reagent10
  • This procedure has been developed by customers for the transient transfection of HEK 293 cells in 75 cm3 flasks using QIAGEN® PolyFect® Transfection Reagent. (qiagen.com)
  • Latest advances in transient transfection protocols using polyethylenimine (PEI) being a transfection reagent possess led to the introduction of cost-effective methods offering yields enough for commercial production of proteins for most preclinical needs. (healthcarecoremeasures.com)
  • One of the most commonly used and economical methods of transient gene expression uses polyethylenimine (PEI) as the transfection reagent (Boussif et al. (healthcarecoremeasures.com)
  • The Effectene Transfection Reagent is intended for molecular biology applications. (qiagen.com)
  • Effectene Transfection Reagent is a nonliposomal lipid reagent for DNA transfection into a broad range of cell types. (qiagen.com)
  • Due to its low cytotoxicity, Effectene Transfection Reagent is highly suitable for transfection of primary cells and many sensitive cell lines. (qiagen.com)
  • Transfection using Effectene Transfection Reagent requires low amounts of DNA and minimal handling. (qiagen.com)
  • In addition, removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening. (qiagen.com)
  • Here we describe transient protein production using QMCF Technology (Icosagen, Estonia) that uses specific suspension-adapted mammalian cell lines (QMCF cells), appropriate QMCF episomal expression vector, a chemically defined animal origin-free, serum-free medium, and Reagent 007™ (Icosagen, Estonia) for efficient delivery of nucleic acids for transfection of mammalian cells in comparison with PEI MAX™ (Transfection Grade Linear Polyethylenimine Hydrochloride, Polysciences). (springer.com)
  • Karro K, Männik T, Männik A, Ustav M (2015) DNA transfer into animal cells using stearylated CPP based transfection reagent. (springer.com)
Polyethylenimine4
  • A scalable transfection procedure using polyethylenimine (PEI) is described for the human embryonic kidney 293 cell line grown in suspension. (canada.ca)
  • Keywords: Transient transfection Polyethylenimine (PEI) Iron (III) citrate Recombinant proteins manifestation Monoclonal antibody Chinese language hamster ovary (CHO) cell range Introduction The usage of monoclonal antibodies for therapeutics offers risen dramatically before few years. (healthcarecoremeasures.com)
  • Longo PA, Kavran JM, Kim M-S, Leahy DJ (2013) Transient mammalian cell transfection with polyethylenimine (PEI). (springer.com)
  • The methodology, an advanced form of DOE that incorporates response surface methods (RSMs), focused on the relative amounts of transfection agent polyethylenimine (PEI) and DNA. (statease.com)
Recombinant1
  • The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. (qiagen.com)
Stable6
  • In transfection, the introduced nucleic acid may be transient, such that it does not replicate, or it may be stable and integrate into the genome of the recipient and replicate when the host genome replicates. (newworldencyclopedia.org)
  • If it is desired that the transfected gene actually remains in the genome of the cell and its daughter cells, a stable transfection must occur. (newworldencyclopedia.org)
  • Transient and stable vector transfection: Pitfalls, off-target effects, artifacts. (nih.gov)
  • These processes include transient transfection, and the creation of stable pools and clonal cell lines. (lonza.com)
  • Transient and stable pool processes are generally used for discovery through early-stage development to enable rapid production and screening of large numbers of protein candidates, while stable clonal cell lines are used for GMP clinical and commercial manufacturing. (lonza.com)
  • This technique has been used to obtain both transient 1 and stable 2 transfections in a wide variety of cell types. (sigmaaldrich.com)
Cells24
  • transfection of suspension-growing cells was more efficient with linear PEI and was not affected by the presence of medium conditioned for 24 h. (canada.ca)
  • Broadly defined, transfection is the process of artificially introducing nucleic acids ( DNA or RNA ) into cells , utilizing means other than viral infection. (newworldencyclopedia.org)
  • Commonly, the term transfection is used in the sense of non-viral introductions of nucleic acid specifically into eukaryotic cells, or even more specifically into animal cells. (newworldencyclopedia.org)
  • Transfection of animal cells typically involves opening transient pores or "holes" in the cell plasma membrane , to allow the uptake of material. (newworldencyclopedia.org)
  • Because the term transformation had another sense in animal cell biology (a genetic change allowing long-term propagation in culture, or acquisition of properties typical of cancer cells), the term transfection acquired, for animal cells, its present meaning of a change in cell properties caused by introduction of nucleic acids by non-viral methods. (newworldencyclopedia.org)
  • 1990) state that the meaning of transfection is "clear and unambiguous" and refers to the "process by which nucleic acid (DNA or RNA) is introduced to cells with the intent to be expressed in the recipient. (newworldencyclopedia.org)
  • The transfection process is different from "infection," which is the term used for a viral method of introducing nucleic acids into cells (Promega). (newworldencyclopedia.org)
  • Transformation of eukaryotic cells in tissue culture is usually called transfection. (newworldencyclopedia.org)
  • Since the DNA introduced in the transfection process is usually not inserted into the nuclear genome, the foreign DNA is lost at the later stage when the cells undergo mitosis . (newworldencyclopedia.org)
  • En face co-immunostaining of the mouse aortic arch revealed a low level of PDCD4 in endothelial cells undergoing pulsatile shear stress. (plos.org)
  • Celetrix cell electroporation large-scale model SLT (catalog No. 11-0104) is a unique CHO electroporation machine for large scale protein transient production in CHO cells. (ushelf.com)
  • the smallest scale (0.2 ml electroporation tube): 4x10E7 CHO cells per electroporation transfection for 8 ml CHO transient production. (ushelf.com)
  • Within this research an evaluation was performed of varied the different parts of a serum-free moderate employed for Chinese language hamster ovary cells where PEI-mediated transient transfection was inhibited. (healthcarecoremeasures.com)
  • 2004). Currently most reports of transient transfection using PEI have used either HEK 293 or Chinese hamster ovary (CHO) cells (Derouazi et al. (healthcarecoremeasures.com)
  • 1995). However CHO cells are used in most manufacturing cell lines and so it is desirable to use CHO cells for transient transfections to maintain the same post-translational modifications that would be found in the clinical or commercial product for applications that are sensitive to these modifications. (healthcarecoremeasures.com)
  • One of the most critical parameters affecting transient transfection efficiency is the medium in which the cells are grown. (healthcarecoremeasures.com)
  • 2008). With this research we have analyzed several different press for his or her compatibility with PEI-mediated transient transfection in CHO cells. (healthcarecoremeasures.com)
  • For transfection experiments, hMSCs were transferred to 24-well plates and seeded at a density of 15 000 cells/well. (edu.pl)
  • After transfection, cells were maintained in culture conditions up to 21 days. (edu.pl)
  • For transient transfection, cells at a confluence of 50% have been washed with serum-free of charge Opti-MEM medium (Lifetime Technologies, Carlsbad, CA, United states), followed by incubation at 37uC in OptiMEM for 4 h with plasmid DNA (2.5 or five mg/ml) and .one% DMRIE-C (Lifetime Systems). (gpr44.com)
  • The exosomes (Exo) from human bone marrow mesenchymal stem cells (hBMSCs) after the transfection of miR mimics/inhibitor/si-PTEN were isolated and used to stimulate M4E and Hep2 cells. (hindawi.com)
  • Data from in vitro translation experiments and transfection of melanoma cells with bicistronic vectors documented that MELOE-3 is exclusively translated by the classical cap-dependent pathway. (oncotarget.com)
  • Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. (sigmaaldrich.com)
  • The procedure stated below is designed for the transfection of CHO cells with 1 µg/µL pSV40-CAT plasmid (diluted in sterile molecular biology grade water). (sigmaaldrich.com)
Electroporation1
Protocols3
Efficiency4
  • Green fluorescent protein (GFP) and human placental secreted alkaline phosphatase (SEAP) were used as reporter genes to monitor transfection efficiency and productivity. (canada.ca)
  • Transfection efficiency was assessed by confocal microscopy using GFP fluorescent signal detection. (edu.pl)
  • Results: MSC pDNA-eGFP transfection results in a dramatically low efficiency, less than 1% of the cell population in each of the tested conditions. (edu.pl)
  • In contrast, mRNA-eGFP transfection resulted in an efficiency exceeding 95% in each of the tested conditions. (edu.pl)
Reagents1
Proteins3
  • Mammalian transient expression systems enable flexible and rapid production of proteins. (springer.com)
  • ACP-tag fusion proteins can be expressed by transient transfection. (neb.com)
  • In this white paper, we describe how GS Discovery® provides optimized transient expression processes to support early stage expression of a range of therapeutic proteins. (lonza.com)
Vectors1
  • Conclusions: The mRNA transfection is a robust, clinically applicable tool for inducing the transient expression of transgenes in hMSCs, which are otherwise difficult to transfect by vectors that do not incorporate into the host genome. (edu.pl)
Invitrogen1
Expression2
  • It is anticipated that, for cell homing to the brain after intra-arterial delivery, the transient expression of integrins should be sufficient for diapedesis to occur. (edu.pl)
  • Since the capacity of MSCs to undergo functional transfection using pDNA is very low, we investigated an mRNA transfection method for the expression of transgenes in MSCs in order to overcome the limitations of the pDNA approach. (edu.pl)
Functional1
  • Transient mismatch repair gene transfection for functional analysis of genetic hMLH1 and hMSH2 variants. (xenbase.org)
Sufficient1
Methods1
Creation1
  • Therefore creation of monoclonal antibodies by transient transfection is now ever more popular for the creation of materials for study and development as well as for in vivo research. (healthcarecoremeasures.com)
Influence1
  • The N/P ratio is well known to influence the rAAV yields obtained via transient transfection. (statease.com)
Mammalian6
  • Devising a simple and cost-effective method for expressing glycoproteins by transient transfection in mammalian cells - where glycosylation processes are native - is therefore highly sought-after and has been an area of intense research activity. (polyplus-sartorius.com)
  • Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. (researchsquare.com)
  • Twenty four to 48 hours after transfection begin selecting mammalian cultures in 600-1,200 µg/ml G418 (geneticin) depending on the cell line. (neb.com)
  • We currently obtain vector batches at high titer and purity using transient transfection of mammalian cells (HEK293) grown in orbital-shaken suspension cultures. (epfl.ch)
  • The plasmids are selectable with Blasticidin in both E. coli and mammalian cells (transient and stable transfection). (invivogen.com)
  • The various transient protein expression systems are yeast, bacteria, insect and mammalian systems. (pharmiweb.com)
Plasmid4
  • Transient transfection allows for the temporary cellular expression of gene products located on an extrinsic plasmid. (polyplus-sartorius.com)
  • Because of differences in transfection efficiencies, an expression plasmid pCMV β-galactosidase will be used to co-transfect as a transfection efficiency control, and luciferease activities will be normalized based on β-galactosidase activity with the chemiluminescent β-gal reporter gene assay kit (Promega). (researchsquare.com)
  • Simplified process flows can also be achieved with producer cells due to elimination of the multi-plasmid transient co-transfection step, observes Peterfay. (biopharminternational.com)
  • It was shown that transient transfection of CHO cells by the Bcl-xL anti-apoptotic protein expressing plasmid prolonged the cell survival rate and protected cells from apoptosis. (iyte.edu.tr)
Stable8
  • CLIP-tag fusion proteins can be expressed by transient or by stable transfection. (neb.com)
  • pSNAP-CaaX has performed well in stable and transient transfection of CHO-K1 and NIH 3T3 cells. (neb.com)
  • Stable Expression pSNAP-CaaX can be transfected as described above for transient transfection or by other standard transfection methods. (neb.com)
  • Transient protein expression is a speedy process when compared to stable integration. (pharmiweb.com)
  • The stable transfection takes months and weeks while transient protein expression is accomplished in days, providing reliable and rapid access to small quantities of proteins for experimentation as well as characterization. (pharmiweb.com)
  • Moreover the market for transient protein expression is expected to increase due to its relatively inexpensive cost when compared to stable transfection also the timeline of expression is rapid. (pharmiweb.com)
  • Cell lines with integrated EJ5 or G4-EJ5 were then produced through stable transfection into human HEK293 cells. (chicagobiomedicalconsortium.org)
  • Each method has its own advantages and disadvantages and may be used for transient or stable transfections. (btxonline.com)
Assays3
  • We investigate transfections in small-scale assays (25 mL) and expand our studies to larger volumes (200 mL). (polyplus-sartorius.com)
  • Octet quantitation assays with anti-His2 biosensors (ForteBio®, cat# 18-5114) that bind the (His)6x tag on gp1 revealed that the highest expression yield was achieved with FectoPRO ® and Booster (Polyplus-transfection®, cat# 116-001). (polyplus-sartorius.com)
  • MUC4 mRNA levels and promoter regulation were studied by reverse transcriptase-PCR and transient transfection assays respectively. (nih.gov)
Protein expression8
  • As 2 O 3 -induced inhibition in myotube formation and muscle-specific protein expression was reversed by transfection with the constitutively active form of Akt . (nih.gov)
  • Transient Protein Expression Market Is Driving With The I. (pharmiweb.com)
  • The Transient Protein Expression market is driving with the increase in development of genetic engineering which has further opened many avenues and possibilities of expression and isolation of proteins in large scale. (pharmiweb.com)
  • Tentatively, Transient Protein Expression market has been segmented on the basis of Product type, Applications, End User and Geography. (pharmiweb.com)
  • The transient protein expression is preferred more for small scale protein expression due to its rapid results. (pharmiweb.com)
  • Regionally, Transient Protein Expression market is segmented into various regions. (pharmiweb.com)
  • North America region is expected to dominate Transient Protein Expression market globally, followed by Europe, due to the rising adoption of technology in academic research centers and both research laboratories. (pharmiweb.com)
  • The growing advancement in the technology with use of cost-effective devices replacing the traditional systems with the newest technologies is increasing the market for Transient Protein Expression devices in the region. (pharmiweb.com)
Assay1
  • Effects on interleukin-8 (IL-8) promoter regulation were determined by transient transfection assay. (cdc.gov)
Serum1
  • The highest glycoprotein yields were observed at six days post-transfection using 293 Freestyle™ Medium Serum Free (Thermo Fisher Scientific®, cat# 12338026) together with FectoPRO® and Booster. (polyplus-sartorius.com)
Commercially available1
  • Moreover one of the major factor driving the improvement of the market includes increasing public-private funding for life science, research and presently commercially available very high-throughput devices for transient protein expressions. (pharmiweb.com)
Yields1
  • The N/P ratio is well known to influence the rAAV yields obtained via transient transfection. (statease.com)
Investigate2
  • We also investigate the impact of performing transient transfection at various cell densities. (polyplus-sartorius.com)
  • At the time, we were performing transient transfections to investigate those seven base pairs in the dynorphin promoter. (nih.gov)
Clinical2
  • In the transient transfection method, full-length HBV genomes from clinical isolates are amplified and cloned. (medscape.com)
  • The most commonly used platform for generating LVVs in the R&D and clinical space relies on transfection of four individual plasmids. (biopharminternational.com)
Genes1
  • Transient transfection analysis of SULT1E1 promoter-luciferase reporter genes revealed that phosphorylated PXR and RORa repressed SULT1E1 promoter activity. (nih.gov)
Analysis1
  • Protein abundance, as determined by Western blot analysis following transient transfection, is not altered. (nih.gov)
Amounts1
  • The methodology, an advanced form of DOE that incorporates response surface methods (RSMs), focused on the relative amounts of transfection agent polyethylenimine (PEI) and DNA. (statease.com)
Time1
Culture1
  • We observed that the culture with a transfection ratio of 0.5:0.5:1.5 DNA:FectoPRO®:Booster yielded the highest amount of gp1, at approximately 3-8 µg/mL six days post-transfection (Fig. 1C). (polyplus-sartorius.com)
Interest1
  • Transfection is simply getting DNA (or other molecules of interest) into a cell, while keeping that cell alive. (btxonline.com)
Manufacturers1
  • Moreover, the manufacturers of Transient Protein Expressions are mainly focusing on strengthening and increasing the core competencies of their product portfolio. (pharmiweb.com)