Gene Transferin vitroandin vivowith Epstein-Barr Virus-Based Episomal Vector Results in Markedly High Transient Expression in Rodent Cells

Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart

“…The specimens were then stained with 6 ], 60 mM Na 2 HPO 4 , 40 mM NaH 2 PO 4 , and 0.1% Triton X-100). After 12 h of incubation at 37°C, the reaction was terminated by replacing the solution with PBS [33].…”

Highly efficient gene transfer into murine liver achieved by intravenous administration of naked Epstein–Barr virus (EBV)-based plasmid vectors

“…By our method, maximum expression of luciferase reached 2 g/mg protein (Figure 2a), indicating that the Although an oriP-bearing plasmid does not replicate in rodent cells, 12,31 we have previously demonstrated that a significantly high expression of a transgene can be achieved in murine and rat cells by using EBV-based plasmid vectors. 29 This may be attributed to EBNA1 functions other than DNA replication, ie nuclear transfer of the plasmid, binding of the plasmid to the nuclear matrix, and transcriptional enhancement. We have recently found that these functions actually work in rodent cells, at least in some in vitro experimental systems (Kishida et al, unpublished observations).…”

“…On the other hand, we have recently shown that naked EBV-based plasmid DNA was quite effective in transducing cardiac muscle. 22,29 When an EBV-based plasmid containing the ␤ 2 -adrenergic receptor gene was injected into the myocardium of cardiomyopathic hamsters, a significant elevation in cardiac output was obtained, suggesting a novel gene therapy strategy against heart failure. 22 The 'hydrodynamics-based procedure' may become an attractive approach for gene therapy of various diseases as well as functional, in vivo analysis of genes of interest.…”

“…[24][25][26][27] In some of these studies, the EBV-based plasmid vectors were combined with cationic liposomes, 25,28 cationic polymers, 23,[25][26][27]29 electroporation 21 or gene gun, 30 while in the other studies, naked EBV-based plasmid was injected into the cardiac muscle. 22,29 In the present study, we transferred the naked EBV-based plasmid vectors to the liver through the intravascular route.EBV-based and conventional plasmid vectors encoding the luciferase or ␤-gal genes as markers (Figure 1, upper panels) were constructed and injected intravenously into the tail vein of mice as described. 9 The injection with pG.GL3, a conventional luciferase-expression vector,…”

“…We have applied the EBV-plasmids to various preclinical studies of gene therapy, including those for hereditary, 21 chronic, 22,23 as well as malignant diseases. [24][25][26][27] In some of these studies, the EBV-based plasmid vectors were combined with cationic liposomes, 25,28 cationic polymers, 23,[25][26][27]29 electroporation 21 or gene gun, 30 while in the other studies, naked EBV-based plasmid was injected into the cardiac muscle. 22,29 In the present study, we transferred the naked EBV-based plasmid vectors to the liver through the intravascular route.…”