Extremely efficient gene transfection into lympho-hematopoietic cell lines by Epstein-Barr virus-based vectors

We have estimated the efficiency of Epstein-Barr virus (EBV)-based vectors in transfecting genes into cell lines of lympho-hematopoietic lineages. The transfection efficiency was estimated both at transient and stable phases, in terms of expression of a marker gene and acquisition of drug resistance, respectively. Plasmid vectors carrying EBV oriP (replication origin of plasmid), EBNA (EBV nuclear antigen)-1 and as the marker genes, murine CD8 alpha cDNA and neoR (neomycin resistant) genes were transfected into various cell lines by electroporation. When cell lines constitutively expressing EBNA-1 were transduced, virtually all the cells expressed CD8 alpha on day 3 and acquired G418 resistance thereafter. In the case of K562 cells, which do not express EBNA-1, approximately 40% of cells expressed the marker gene product on day 3 posttransfection, and 30% of cells became stable transfectants. These data suggest a broader application of the EBV vector system in basic immunology and medicine.