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Transient Mammalian Cell Transfection with Polyethylenimine (PEI)

Standard protein expression systems, such as E. coli, often fail to produce folded, monodisperse, or functional eukaryotic proteins (see Small-scale Expression of Proteins in E. coli). The expression of these proteins is greatly benefited by using a eukaryotic system, such as mammalian cells, that c... Full description

Journal Title: Methods in Enzymology 2013, Vol.529, p.227-240
Main Author: Longo, Patti A
Other Authors: Kavran, Jennifer M , Kim, Min-Sung , Leahy, Daniel J
Format: Electronic Article Electronic Article
Language: English
Subjects:
Publisher: United States: Elsevier Science & Technology
ID: ISSN: 0076-6879
Link: https://www.ncbi.nlm.nih.gov/pubmed/24011049
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title: Transient Mammalian Cell Transfection with Polyethylenimine (PEI)
format: Article
creator:
  • Longo, Patti A
  • Kavran, Jennifer M
  • Kim, Min-Sung
  • Leahy, Daniel J
subjects:
  • Adherent cells
  • Animals
  • Article
  • Cell Culture Techniques
  • Cell Line - cytology
  • Cells (Biology)
  • Escherichia coli
  • Gene Expression
  • Harvest cells
  • Humans
  • Infection
  • Mammalian cell transfection
  • Polyethyleneimine - pharmacology
  • Polyethylenimine (PEI)
  • Proteins
  • Proteins - genetics
  • Protocol
  • Transfect cells
  • Transfection - methods
  • Transient transfections
ispartof: Methods in Enzymology, 2013, Vol.529, p.227-240
description: Standard protein expression systems, such as E. coli, often fail to produce folded, monodisperse, or functional eukaryotic proteins (see Small-scale Expression of Proteins in E. coli). The expression of these proteins is greatly benefited by using a eukaryotic system, such as mammalian cells, that contains the appropriate folding and posttranslational machinery. Here, we describe methods for both small- and large-scale transient expression in mammalian cells using polyethylenimine (PEI). We find this procedure to be more cost-effective and quicker than the more traditional route of generating stable cell lines. First, optimal transfection conditions are determined on a small-scale, using adherent cells. These conditions are then translated for use in large-scale suspension cultures. For further details on generating stable cell lines please (see Rapid creation of stable mammalian cell lines for regulated expression of proteins using the Gateway® Recombination Cloning Technology and Flp-In T-REx® lines or Generating mammalian stable cell lines by electroporation).
language: eng
source:
identifier: ISSN: 0076-6879
fulltext: no_fulltext
issn:
  • 0076-6879
  • 1557-7988
url: Link


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descriptionStandard protein expression systems, such as E. coli, often fail to produce folded, monodisperse, or functional eukaryotic proteins (see Small-scale Expression of Proteins in E. coli). The expression of these proteins is greatly benefited by using a eukaryotic system, such as mammalian cells, that contains the appropriate folding and posttranslational machinery. Here, we describe methods for both small- and large-scale transient expression in mammalian cells using polyethylenimine (PEI). We find this procedure to be more cost-effective and quicker than the more traditional route of generating stable cell lines. First, optimal transfection conditions are determined on a small-scale, using adherent cells. These conditions are then translated for use in large-scale suspension cultures. For further details on generating stable cell lines please (see Rapid creation of stable mammalian cell lines for regulated expression of proteins using the Gateway® Recombination Cloning Technology and Flp-In T-REx® lines or Generating mammalian stable cell lines by electroporation).
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languageeng
publisherUnited States: Elsevier Science & Technology
subjectAdherent cells ; Animals ; Article ; Cell Culture Techniques ; Cell Line - cytology ; Cells (Biology) ; Escherichia coli ; Gene Expression ; Harvest cells ; Humans ; Infection ; Mammalian cell transfection ; Polyethyleneimine - pharmacology ; Polyethylenimine (PEI) ; Proteins ; Proteins - genetics ; Protocol ; Transfect cells ; Transfection - methods ; Transient transfections
ispartofMethods in Enzymology, 2013, Vol.529, p.227-240
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descriptionStandard protein expression systems, such as E. coli, often fail to produce folded, monodisperse, or functional eukaryotic proteins (see Small-scale Expression of Proteins in E. coli). The expression of these proteins is greatly benefited by using a eukaryotic system, such as mammalian cells, that contains the appropriate folding and posttranslational machinery. Here, we describe methods for both small- and large-scale transient expression in mammalian cells using polyethylenimine (PEI). We find this procedure to be more cost-effective and quicker than the more traditional route of generating stable cell lines. First, optimal transfection conditions are determined on a small-scale, using adherent cells. These conditions are then translated for use in large-scale suspension cultures. For further details on generating stable cell lines please (see Rapid creation of stable mammalian cell lines for regulated expression of proteins using the Gateway® Recombination Cloning Technology and Flp-In T-REx® lines or Generating mammalian stable cell lines by electroporation).
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abstractStandard protein expression systems, such as E. coli, often fail to produce folded, monodisperse, or functional eukaryotic proteins (see Small-scale Expression of Proteins in E. coli). The expression of these proteins is greatly benefited by using a eukaryotic system, such as mammalian cells, that contains the appropriate folding and posttranslational machinery. Here, we describe methods for both small- and large-scale transient expression in mammalian cells using polyethylenimine (PEI). We find this procedure to be more cost-effective and quicker than the more traditional route of generating stable cell lines. First, optimal transfection conditions are determined on a small-scale, using adherent cells. These conditions are then translated for use in large-scale suspension cultures. For further details on generating stable cell lines please (see Rapid creation of stable mammalian cell lines for regulated expression of proteins using the Gateway® Recombination Cloning Technology and Flp-In T-REx® lines or Generating mammalian stable cell lines by electroporation).
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