title: Transient mammalian cell transfection with polyethylenimine (PEI).
abstract: 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).
authors.affiliation: Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Patti A
authors.initials: PA
authors.lastname: Longo
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Kavran
authors.forename: Min-Sung
authors.initials: MS
authors.lastname: Kim
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Leahy
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line / Q000166:cytology*; D004926:Escherichia coli; D015870:Gene Expression; D006801:Humans; D011094:Polyethyleneimine / Q000494:pharmacology*; D011506:Proteins / Q000235:genetics*; D014162:Transfection / Q000379:methods*
chemical_list: D011506:Proteins; D011094:Polyethyleneimine
keywords: Adherent cells; Harvest cells; Mammalian cell transfection; Polyethylenimine (PEI); Protocol; Transfect cells; Transient transfections