Detection of plasmid DNA vectors following gene transfer to the murine airways

Pringle, Ian A.; Raman, S; Sharp, W W; Cheng, Seng H.; Hyde, Stephen C.; Gill, Deborah R.

doi:10.1038/sj.gt.3302518

Cited by 40 publications

(30 citation statements)

References 43 publications

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“…naked plasmid DNA or DNA/liposomes) often lead to transient gene expression when delivered to the mouse lung, despite the persistence of intact plasmid DNA. [40][41][42] Strong viral promoters such as the cytomegalovirus (CMV IE) promoter can become attenuated in the lung, resulting in silencing of transgene expression after only a few days. 41,43 However, as we report here, CMVmediated reporter gene expression from rAAV5/5 is persistent under similar conditions.…”

Section: Discussionmentioning

confidence: 99%

Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways

et al. 2006

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Section: Discussionmentioning

confidence: 99%

Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways

et al. 2006

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“…4 Recently, it was reported that the transience of transgene expression was due to a decrease in the transcription efficiency. [5][6][7][8][9][10] The expression from transgenes introduced by a rapid, high-volume injection (hydrodynamics-based injection) is also generally transient, [5][6][7]9 although prolonged transgene expression was reported in some cases. 6,7,[11][12][13][14] One possible factor involved in the 'silencing' of the exogenous DNA is the methylation of the 5-position of cytosine in the CpG motifs.…”

Section: Introductionmentioning

confidence: 99%

“…However, we and others found that the silencing of the exogenous DNA proceeded without the DNA methylation. 8,9 Thus, the involvement of the CpG methylation in the transgene silencing is still debatable.…”

Section: Introductionmentioning

confidence: 99%

Transient activation of transgene expression by hydrodynamics-based injection may cause rapid decrease in plasmid DNA expression

et al. 2007

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The intranuclear disposition of exogenous DNA is quite important for the therapeutic effects of the administered DNA. The expression efficiency from one copy of exogenous DNA delivered by hydrodynamics-based injection dramatically decreases over time, and this 'silencing' occurs without CpG methylation. In this study, naked luciferase-plasmid DNA was delivered into mouse liver by hydrodynamics-based injection, and modifications of the histones bound to the plasmid DNA were analyzed by a chromatin immunoprecipitation (ChIP) analysis. In addition, the effects of a second hydrodynamics-based injection on the expression from the plasmid DNA were examined.The ChIP analysis revealed that the modification status of histone H3 remained constant from 4 h to 4 weeks. Surprisingly, the injection of saline without DNA enhanced the luciferase expression from the preexisting DNA administered 4 and 14 days previously. Our results suggest that histone modification plays no role in the silencing. Instead, our data suggest that the transgene expression is activated by the hydrodynamics-based injection manipulation, and that the return from the activated status causes the silencing.

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“…They found that the methylation status did not change in mouse organs. 22,23) Regarding histone modifications, we did not obtain any evidence that such modifications were involved in the silencing.…”

Section: Discussionmentioning

confidence: 69%

Effects of Endogenous Proteins and microRNA Target Sequence in a Positive Feedback System

Kanda

Togashi

Harashima

et al. 2012

Biological & Pharmaceutical Bulletin

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A positive feedback system, using GAL4-vp16 (a fusion protein of yeast GAL4 and herpes simplex virus vp16) as an activator and firefly luciferase as a reporter, maintained luciferase expression for 7 d in mice. However, the luciferase expression decreased after 7 d, and this phenomenon could be caused by immunoreactions against these exogenous proteins. This hypothesis was examined by the following three strategies, designed to avoid the putative immunoreactions: (i) use of the endogenous secreted alkaline phosphatase (SEAP) protein as a reporter, (ii) replacement of vp16 with endogenous transcription factors, and (iii) insertion of the target sequence of microRNA expressed in cells of hematopoietic origin, to suppress GAL4-vp16 expression in antigen-presenting cells. The results obtained in this study suggested that silencing would be induced by mechanism(s) besides immunoreactions against reporter and activator proteins.

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Detection of plasmid DNA vectors following gene transfer to the murine airways

Cited by 40 publications

References 43 publications

Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways

Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways

Transient activation of transgene expression by hydrodynamics-based injection may cause rapid decrease in plasmid DNA expression

Effects of Endogenous Proteins and microRNA Target Sequence in a Positive Feedback System

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