title: Nanosecond electric pulse effects on gene expression.
abstract: Gene electrotransfection using micro- or millisecond electric pulses is a well-established method for safe gene transfer. For efficient transfection, plasmid DNA has to reach the nucleus. Shorter, high-intensity nanosecond electric pulses (nsEPs) affect internal cell membranes and may contribute to an increased uptake of plasmid by the nucleus. In our study, nsEPs were applied to Chinese hamster ovary (CHO) cells after classical gene electrotransfer, using micro- or millisecond pulses with a plasmid coding the green fluorescent protein (GFP). Time gaps between classical gene electrotransfer and nsEPs were varied (0.5, 2, 6 and 24 h) and three different nsEP parameters were used: 18 ns-10 kV/cm, 10 ns-40 kV/cm and 15 ns-60 kV/cm. Results analyzed by either fluorescence microscopy or flow cytometry showed that neither the percentage of electrotransfected cells nor the amount of GFP expressed was increased by nsEP. All nsEP parameters also had no effects on GFP fluorescence intensity of human colorectal tumor cells (HCT-116) with constitutive expression of GFP. We thus conclude that nsEPs have no major contribution to gene electrotransfer in CHO cells and no effect on constitutive GFP expression in HCT-116 cells.
authors.affiliation: CNRS, IPBS - UMR 5089, 205 route de Narbonne, BP 64182, 31077, Toulouse Cedex 4, France.
authors.forename: Louise
authors.initials: L
authors.lastname: Chopinet
authors.forename: Tina
authors.initials: T
authors.lastname: Batista-Napotnik
authors.forename: Audrey
authors.initials: A
authors.lastname: Montigny
authors.forename: Matej
authors.initials: M
authors.lastname: Rebersek
authors.forename: Justin
authors.initials: J
authors.lastname: Teissié
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Rols
authors.forename: Damijan
authors.initials: D
authors.lastname: Miklavčič
mesh_terms: D000818:Animals; D016466:CHO Cells; D002463:Cell Membrane Permeability; D006224:Cricetinae; D003412:Cricetulus; D004558:Electric Stimulation; D018274:Electroporation; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D045325:HCT116 Cells; D006801:Humans; D009685:Nuclear Envelope / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins