An EBV‐based plasmid can replicate and maintain in stem cells

Abstract

Viral vectors have a wide range of applications in biology, particularly in gene therapy. Based on their integration capacity, viral vectors are classified as either integrating or non-integrating vectors. Although integrating vectors, such as lentivectors, have the ability to direct prolonged expression of exogenous genes, manipulation of the host genome is an inappropriate feature of these gene delivery tools. Non-integrating vectors, such as episomal replicating plasmids, can replicate and persist in host cells for long periods without any chromosomal interruption. These advantages made them good tools for gene induction purposes in gene therapy and basic studies. Due to the necessity of gene induction in stem cells for study of mammalian development and targeted differentiation, the use of integrating vectors for prolonged expression of genes of interest has been developed. Application of replicating plasmids can overcome some drawbacks associated with integrating vectors, although replication and maintenance of these plasmids can differ between cell types. Previously, it has been shown that such plasmids can be maintained in human embryonic stem cells for more than one month, but the rate of the plasmid replication during the host cell cycle has not been elucidated. In the present study, we showed that an EBV-based plasmid can replicate simultaneously with host in pluripotent and multipotent human and mouse stem cells and can be sustained for long time periods in dividing cells.

Plasmid-Based Generation of Induced Neural Stem Cells from Adult Human Fibroblasts

“…Another study employing vectors containing the oriP/EBNA1 system for direct reprogramming could not detect ongoing expression of reprogramming factors ( Kim et al, 2016 ). However, other studies evaluating the feasibility of oriP/EBNA1 as an expression system in human and murine stem cells demonstrated a prolonged expression ( Ali Hosseini Rad et al, 2015 ).…”

Advances in Non-Viral DNA Vectors for Gene Therapy

“…Epstein–Barr virus (EBV) is a herpes virus that replicates its genome episomally in host cells upon latent infection [ 125 ] by expressing Epstein–Barr nuclear antigen 1 (EBNA-1), which in turn recognizes the oriP site and initiates replication [ 125 ]. This system was exploited by encoding oriP and EBNA-1 onto a DNA vector for the long-term replication of plasmids in mesenchymal stem cells [ 126 ] and human fibroblasts [ 127 ]. Other viruses, such as SV40 and papillomaviruses, have similar systems with trans -acting elements that allow them to replicate in mammalian cells [ 128 ].…”