Transgene expression enhancement in T-lymphoma cell lines.

P. RUYBAL; M.J. GRAVISACO; V. BARCALA; A. ESCALADA; G.A. CREMASCHI; O. TABOGA; C. WALDNER; C. MONGINI

INTERNATIONAL IMMUNOPHARMACOLOGY

Elsevier Science

Lugar: USA; Año: 2005 vol. 5 p. 1685 - 1692

1567-5769

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.