Chromatin-binding regions of EBNA1 protein facilitate the enhanced transfection of Epstein-Barr virus-based vectors.

摘要: Epstein-Barr virus (EBV)-based vectors can stably maintain large genomic fragments in mammalian cells, offering great potential for the treatment/correction of many acquired and inherited disorders. Numerous studies report marked increases transfection efficiency EBV-based after delivery into cell lines constitutively expressing nuclear antigen-1 (EBNA1), compared with cells not EBNA1. We employ a novel strategy, involving mRNA encoding EBNA1, to transiently express EBNA1 protein human cells. Subsequently we show that 21-kb EBVbased vector is improved significantly when codelivered Similar were observed plasmid also investigate mechanism by which facilitates vectors, using modified versions protein. Previous suggest DNA-binding domain (DBD), together localization signal (NLS), may enhance EBV plasmids facilitating their transport. demonstrate an derivative comprising only NLS DBD does facilitate vectors. However, devoid functional but retaining chromatin-binding regions, domains A B, enhances up 10-fold. Moreover, variant two copies fused DNA even greater extent than wild-type therefore propose EBNA1-mediated dependent on presence chromatin- binding regions DBD, NLS.