Up to 100-Fold Increase of Apparent Gene Expression in the Presence of Epstein-Barr Virus
            <i>oriP</i>
            Sequences and EBNA1: Implications of the Nuclear Import of Plasmids

Längle-Rouault, Francoise; Patzel, Volker; Benavente, Annie; Taillez, Martine; Silvestre, Nathalie; Bompard, Albine; Sczakiel, Georg; Jacobs, Eric J.; Rittner, Karola

doi:10.1128/jvi.72.7.6181-6185.1998

Cited by 121 publications

(34 citation statements)

References 29 publications

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“…Cytoplasmic injection of these plasmids led to an enhancement of luciferase activity of up to 100-fold. This difference in the factor of activation after nuclear or cytoplasmic injection could be ascribed to an increased transport of plasmids carrying oriP from the cytoplasm to the nucleus in the presence of Epstein-Barr nuclear antigen 1 (EBNA1) (Langle-Rouault et al, 1998;Kaneda et al, 2000;Tu et al, 2000). These data suggest the possibility to substantially increase the expression of a gene under the control of a strong constitutive promoter in the presence of oriP sequences and EBNA1.…”

Section: Association Of Dna With Virus-derived Proteins or Peptidesmentioning

confidence: 99%

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Hebert

2003

Biology of the Cell

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Several vectors have been developed in order to target genes to specific cells. Virus-based vectors lead to a high transfection efficiency in vitro, but display important disadvantages such as pathological risks, which they expose to patients. Plasmid-associated chemical vectors lack these disadvantages, but allow only a very low efficiency of transgene expression. Most of the non-viral-based gene transfer techniques developed until now mainly focused their efforts to overcome the problem of DNA entry into the cell. Some recent works, however, have begun to investigate the nucleus entry problem and suggest that the trafficking of DNA from cytosol to the nucleus may be improved by using the nuclear localization signal (NLS) found in some nuclear proteins. If the vector contains one or several NLS, either as covalently or non-covalently DNA-linked peptides, a competition may take place between the rate of dissociation of the DNA-vector complexes and the rate of loading of the complexes to the NLS-mediated nucleus importation machinery. This equilibrium may be displaced towards the importation pathway by the use of NLS-bearing proteins instead of peptides. The possibility of recruiting normal endogenous cellular pathways of nuclear uptake to promote entry of exogenously applied DNA through the nuclear pore complex would, thus, seem promising. Nevertheless, attempts to improve the transport of DNA to the nucleus through the use of NLSs have achieved limited success. Although these systems show improved transgene expression, little is known about how they function in transfected cells, and the optimal formulation for gene expression is yet to be determined.

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Section: Association Of Dna With Virus-derived Proteins or Peptidesmentioning

confidence: 99%

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Hebert

2003

Biology of the Cell

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“…A system for in vivo study of the involvement of chromatin remodelling during transcription elongation was generated using pREP4-Luc episomal reporter vector. pREP4 contains Epstein-Barr virus replication origin and encodes nuclear antigen 1 (Langle-Rouault et al, 1998). This episomal vector has been shown previously to form a correct nucleosomal chromatin structure on transfection (van der Vlag et al, 2000).…”

Section: Description Of the Reporter Systemmentioning

confidence: 99%

BRG1 helps RNA polymerase II to overcome a nucleosomal barrier during elongation, in vivo

Subtil‐Rodríguez

Reyes

2010

EMBO Reports

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RNA polymerase II (RNAPII) transcribes genes in a chromatin context. We have designed a system to investigate the role of chromatin remodelling during elongation in vivo, which involves inserting a strong nucleosome-positioning sequence between a promoter and a reporter gene. Our data indicate that a nucleosome positioned in the body of a transcription unit impairs RNAPII progression, provokes RNAPII accumulation upstream to the positioned nucleosome and reduces transcription. By using this system, we show that BRG1, the enzymatic motor of the SWI-SNF chromatin-remodelling complex, is recruited to the positioned nucleosome in a transcription elongation-dependent manner and facilitates traversal of the nucleosome by RNAPII.

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“…The higher gene expression seen in the presence of EBV components possibly represents a stimulation of hFIX transcription. The EBNA1-oriP interaction has been documented to have transcriptional enhancer activity in numerous studies (10)(11)(12)(13)(14)(15)(16)(17). However, the mechanism by which this occurs is not completely understood.…”

Section: Discussionmentioning

confidence: 99%

“…In cases where the host cells are dividing, the special replication and nuclear retention properties of EBV vectors are beneficial and give a far better outcome for gene expression than conventional plasmid DNA (8,9). Even when DNA replication is not expected, several studies have suggested that gene expression is enhanced in the presence of these EBV sequences (10)(11)(12)(13)(14)(15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Epstein‐Barr Virus Vectors Provide Prolonged Robust Factor IX Expression in Mice

Sclimenti

Neviaser

Baba

et al. 2003

Biotechnology Progress

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We demonstrate that vectors incorporating components from Epstein-Barr virus (EBV) for retention and from human genomic DNA for replication greatly enhance the level and duration of marker gene expression in dividing cultured cells. The same types of vectors were tested in vivo by high-pressure tail vein injection of naked DNA in mice, resulting in liver delivery and expression. The therapeutic gene was a human factor IX (hFIX) minigene comprising genomically derived 5', 3', and intronic sequences that provided relatively good gene expression in vivo. We demonstrated that addition of the EBV EBNA1 gene and its family of repeats binding sites provided a 10- to 100-fold increase in prolonged hFIX expression in mouse liver. A single 25-microg dose of vector DNA generated normal (>5 microg/mL) levels of hFIX throughout the 8 month duration of the experiment. Vector DNA with or without the EBV sequences was retained in liver cells, and vector replication was not a factor in these nondividing liver cells. Instead, it appears that enhancement of stable hFIX expression by the EBV components was responsible for the increased level and duration of therapeutic gene expression. The EBV sequences also significantly enhanced stable expression of a vector carrying the full genomic hFIX gene delivered to mouse liver. These results underline the crucial importance of appropriate gene expression signals on gene therapy vectors and the utility of EBV sequences in particular for increasing stable gene expression.

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Up to 100-Fold Increase of Apparent Gene Expression in the Presence of Epstein-Barr Virus oriP Sequences and EBNA1: Implications of the Nuclear Import of Plasmids

Cited by 121 publications

References 29 publications

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

BRG1 helps RNA polymerase II to overcome a nucleosomal barrier during elongation, in vivo

Epstein‐Barr Virus Vectors Provide Prolonged Robust Factor IX Expression in Mice

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