A novel micro-linear vector for in vitro and in vivo gene delivery and its application for EBV positive tumors

PLoS One. 2012;7(10):e47159. doi: 10.1371/journal.pone.0047159. Epub 2012 Oct 15.

Abstract

Background: The gene delivery vector for DNA-based therapy should ensure its transfection efficiency and safety for clinical application. The Micro-Linear vector (MiLV) was developed to improve the limitations of traditional vectors such as viral vectors and plasmids.

Methods: The MiLV which contained only the gene expression cassette was amplified by polymerase chain reaction (PCR). Its cytotoxicity, transfection efficiency in vitro and in vivo, duration of expression, pro-inflammatory responses and potential application for Epstein-Barr virus (EBV) positive tumors were evaluated.

Results: Transfection efficiency for exogenous genes transferred by MiLV was at least comparable with or even greater than their corresponding plasmids in eukaryotic cell lines. MiLV elevated the expression and prolonged the duration of genes in vitro and in vivo when compared with that of the plasmid. The in vivo pro-inflammatory response of MiLV group was lower than that of the plasmid group. The MEKK1 gene transferred by MiLV significantly elevated the sensitivity of B95-8 cells and transplanted tumor to the treatment of Ganciclovir (GCV) and sodium butyrate (NaB).

Conclusions: The present study provides a safer, more efficient and stable MiLV gene delivery vector than plasmid. These advantages encourage further development and the preferential use of this novel vector type for clinical gene therapy studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cell Line, Tumor
  • Cytokines / blood
  • Cytokines / immunology
  • Genetic Therapy
  • Genetic Vectors / genetics*
  • Genetic Vectors / immunology
  • Genetic Vectors / therapeutic use*
  • Herpesvirus 4, Human / isolation & purification*
  • Humans
  • Immunity, Cellular
  • MAP Kinase Kinase Kinase 1 / genetics*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Neoplasms / genetics
  • Neoplasms / therapy*
  • Neoplasms / virology*
  • Plasmids / genetics
  • Transfection*

Substances

  • Cytokines
  • MAP Kinase Kinase Kinase 1

Grants and funding

This work was supported by Supported by the National Natural Science Foundation of China (Grant No. 31101071), National Basic Research Program of China (973 Program, No. 2011CB9358003), and National Natural Science Foundation of China (No. 30873032and 81071712). The research was supported, in part, by a Discovery Grant from the National Science and Engineering Research Council of Canada (Project # 326415-07). JPG was supported by the Canada Research Chair program, an at large Chair Professorship at the Department of Biology and Chemistry and State Key Laboratory in Marine Pollution, City University of Hong Kong, The Einstein Professor Program of the Chinese Academy of Sciences and the Visiting Professor Program of King Saud University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.