Maintenance of Epstein-Barr virus-derived episomal vectors in the murine Sp2/0 myeloma cell line is dependent upon exogenous expression of human EBP2.
Vectors carrying the origin of replication (oriP) and driving expression of the EBNA-1 protein from Epstein-Barr virus (EBV) replicate as extrachromosomal episomes in human cells. Whether these vectors can be maintained as episomes in murine cells is still controversial. Here we demonstrate that EBNA-1 expression alone was unable to maintain episomal expression of an EBV-based vector in the murine Sp2/0 cell line. However, we were able to obtain long-term episome maintenance in Sp2/0 cells after exogenously expressing human EBP2 by genetic engineering. Our results provide further evidence for the fundamental role of human EBP2 in episomal maintenance of EBV-based vectors. Moreover, we demonstrate that EBV-based vectors can be successfully used in cells presumably incompetent for episomal maintenance.
Habel
Marie-Eve
ME
Héma-Québec, Recherche et développement, Pavillion Héma-Québec, 1009 Route du Vallon, Sainte Foy, QC G1V 4C3, Canada.
Drouin
Mathieu
M
Jung
Daniel
D
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D004261:DNA Replication / Q000235:genetics*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D009101:Multiple Myeloma / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D016601:RNA-Binding Proteins; D018741:Replication Origin / Q000235:genetics*
D002352:Carrier Proteins; C432687:EBNA1BP2 protein, human; C499009:Ebna1bp2 protein, mouse; D019309:Epstein-Barr Virus Nuclear Antigens; D047908:Intracellular Signaling Peptides and Proteins; D016601:RNA-Binding Proteins; C417029:EBV-encoded nuclear antigen 1