Transcriptionally active drugs improve adenovirus vector performance in vitro and in vivo

Gaetano, Carlo; Catalano, Alfonso; Palumbo, Roberta; Illi, Barbara; Orlando, Giuseppe; Ventoruzzo, G; Serino, F.; Capogrossi, Maurizio C.

doi:10.1038/sj.gt.3301296

Cited by 46 publications

(31 citation statements)

References 24 publications

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“…15 In mouse eyes in which expression of luciferase had declined after intravitreous injection of an adenoviral vector containing a luciferase reporter gene driven by a CMV promoter, reactivation of expression occurred after systemic administration of tRA. 16 If transcriptional silencing of the CMV promoter plays a substantial role in the transient expression profile experienced in many studies investigating adenoviralmediated gene transfer, then use of other promoters should be tested.…”

Section: Discussionmentioning

confidence: 99%

Sustained expression after nonviral ocular gene transfer using mammalian promoters

et al. 2006

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The CMV promoter drives high transgene expression and is one of the most commonly used promoters for gene transfer. Tissue-specific mammalian promoters provide an alternative, and it would be useful to have a system to directly compare them to viral promoters free from potential confounding vector-related effects. In this study, we describe how electroporation after subretinal injection of plasmid DNA can be used to perform comparative quantitative analysis of promoter activities. Luciferase assay of eyecup homogenates was carried out after coinjection/electroporation of pGL2, a plasmid containing the promoter fragment of interest coupled to the firefly luciferase gene, and pRL-CMV, a plasmid containing the CMV promoter coupled to the Renilla luciferase gene for normalization. This technique was used to compare activity of different fragments of the 5 0 -upstream region of the vitelliform macular dystrophy 2 ( VMD2) gene, which is selectively expressed in the retinal pigmented epithelial (RPE) cells, and results indicated positive regulatory elements between À104 and À154 bp and between À424 and À585 bp. Addition of a fragment from intron 1 reduced the activity of the À585/+38 bp fragment by 75%. Deletion analysis implicated a 342 bp region near the 5 0 -end of intron 1 in the repression. Results of transient transfections in two cell lines that constitutively express VMD2 were similar, and results in transgenic mice were consistent, providing validation for promoter analysis by in vivo electroporation. We then explored the time course of expression of the À585/+38 VMD2 promoter fragment and found that compared to cassettes driven by CMV or SV40 promoters, which showed peak luciferase activity on day 2 followed by a rapid decrease in activity, the VMD2 promoter fragment showed lower activity initially, but the activity was sustained for up to 56 days (longest time point measured). A promoter fragment from another RPE-specific gene, Rpe65, showed a similar pattern of sustained expression for at least 112 days. These data indicate that nonviral gene transfer can be used to quantitatively evaluate the activity of promoter fragments independent of influence from viral vectors. A potentially important finding using this new technique is the demonstration that relatively sustained passenger gene expression can be achieved with nonviral gene transfer using mammalian rather than viral promoters. Gene Therapy (2006) 13, 798-804.

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Section: Discussionmentioning

confidence: 99%

Sustained expression after nonviral ocular gene transfer using mammalian promoters

et al. 2006

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“…This is consistent with previous reports using this construct. 25,26 Adenovirus expressing the luciferase cDNA (Ad-CMV-Luc) was obtained from the Vector Core facility, Molecular Medicine Institute, University of Pittsburgh. At least 1000 nuclei in the muscle border regions of at least two muscle samples in each case were counted for each condition.…”

Section: Adenovirusmentioning

confidence: 99%

Angiogenesis is confined to the transient period of VEGF expression that follows adenoviral gene delivery to ischemic muscle

Graham

et al. 2005

Gene Ther

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“…122 Taking a different approach to this problem in AdVmediated gene transfer, it has been shown that transcriptionally active drugs such as retinoic acid (RA) in combination with histone deacetylase inhibitor trichostatin A (TSA) can enhance and prolong transgene expression in cell lines and skeletal muscle up to seven-fold over controls. 123 This effect may be mediated, in part, by the direct activation of RA receptors on the CMV promoter which contains repeated RA responsive element (RARA). Perhaps more importantly, these drugs may reactivate expression of the transgene via remodelling of chromatin.…”

Section: Local Ischemia Vasodilation and High-pressure Injectionmentioning

confidence: 99%

Non-viral gene delivery in skeletal muscle: a protein factory

Lu¹,

Bou‐Gharios²,

Partridge³

2003

Gene Ther

168

109

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Ever since the publication of the first reports in 1990 using skeletal muscle as a direct target for expressing foreign transgenes, an avalanche of papers has identified a variety of proteins that can be synthesized and correctly processed by skeletal muscle. The impetus to the development of such applications is not only amelioration of muscle diseases, but also a range of therapeutic applications, from immunization to delivery of therapeutic proteins, such as clotting factors and hormones. Although the most efficient way of introducing transgenes into muscle fibres has been by a variety of recombinant viral vectors, there are potential benefits in the use of non-viral vectors. In this review we assess the recent advances in construction and delivery of naked plasmid DNA to skeletal muscle and highlight the options available for further improvements to raise efficiency to therapeutic levels.

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Transcriptionally active drugs improve adenovirus vector performance in vitro and in vivo

Cited by 46 publications

References 24 publications

Sustained expression after nonviral ocular gene transfer using mammalian promoters

Sustained expression after nonviral ocular gene transfer using mammalian promoters

Angiogenesis is confined to the transient period of VEGF expression that follows adenoviral gene delivery to ischemic muscle

Non-viral gene delivery in skeletal muscle: a protein factory

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