title: Transgene expression enhancement in T-lymphoma cell lines.
abstract: In transfection protocols, the expression levels of the transgene is important to define, still is difficult to obtain in certain cell lines such as those derived from T-lymphoma cells. In this study we evaluate transgene expression kinetics in the presence and absence of two well known transcription activators such as phorbol-12-myristate13-acetate (PMA) and Ionomicin (IO). Three murine T lymphoma cell lines (LBC, EL4 and BW5147) were transfected by electroporation using green fluorescent protein (GFP) as a reporter gene and analyzed by flow cytometry. Addition of PMA/IO resulted in a significant increase of the Mean Fluorescence Intensity but not in GFP-positive cell percentages, either in transient or stable transfected LBC and EL4 cells. Remarkable, BW5147 cells showed low GFP induction with a significant increment only in stable transfected cells. Our results demonstrated that CMV promoter activity can be enhanced in transfected lymphoma cells by PMA/IO suggesting that transgene expression levels can be optimized by means of the use of transcription activators.
authors.affiliation: Laboratorio de Inmunología Molecular y Celular, Centro de Estudios Farmacológicos y Botánicos, CEFYBO-CONICET, Buenos Aires, Argentina.
authors.forename: Paula
authors.initials: P
authors.lastname: Ruybal
authors.forename: María José
authors.initials: MJ
authors.lastname: Gravisaco
authors.forename: Virna
authors.initials: V
authors.lastname: Barcala
authors.forename: Ana
authors.initials: A
authors.lastname: Escalada
authors.forename: Graciela
authors.initials: G
authors.lastname: Cremaschi
authors.forename: Oscar
authors.initials: O
authors.lastname: Taboga
authors.forename: Claudia
authors.initials: C
authors.lastname: Waldner
authors.forename: Claudia
authors.initials: C
authors.lastname: Mongini
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D018274:Electroporation; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics; D015759:Ionomycin / Q000494:pharmacology; D016399:Lymphoma, T-Cell; D051379:Mice; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics*
chemical_list: D049452:Green Fluorescent Proteins; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate