Time course of gene expression after plasmid DNA gene transfer to the liver

Herweijer, Hans; Zhang, Guofeng; Субботин, Владимир; Budker, Vladimir G.; Williams, Phillip; Wolff, Jon A.

doi:10.1002/jgm.178

Cited by 132 publications

(102 citation statements)

References 54 publications

(69 reference statements)

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“…Previously, Herweijer et al measured amounts of plasmid DNA and transgene expression after intraportal delivery of DNA to liver [26]. Their results that the expression decreases more rapidly than DNA is consistent with one of our findings, although we also focused on earlier time points than they did.…”

Section: Discussionsupporting

confidence: 92%

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

2006

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The intranuclear disposition of exogenous DNA is highly important for the therapeutic effects of the administrated DNA. Naked luciferase-plasmid DNA was delivered into mouse liver by a hydrodynamics-based injection, and the amounts of intranuclear plasmid DNA, luciferase, and its mRNA were quantitated at various time points. Methylation of the promoter of the luciferase gene was also analyzed. Expression efficiency from one copy of the exogenous DNA dramatically decreased over time, and the DNA was methylated and degraded into fragments. Unexpectedly, methylation of the intact plasmid DNA was low and did not increase over time. Rather, the fragmented DNA was methylated more frequently than the intact plasmid. These results suggest that the CpG methylation and the degradation of exogenous DNA, and its 'silencing', occurred in parallel in the nucleus.

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Section: Discussionsupporting

confidence: 92%

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

2006

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“…The pDNA delivered to skeletal muscles of rodents or primates is retained in muscle fibers and expresses the encoded gene product for many months, if not years (Wolff et al, 1992;Danko et al, 1993Danko et al, , 1997Zhang et al, 2004Zhang et al, , 2010Sebestyen et al, 2007). Long-term expression of foreign genes can be obtained from naked pDNA without chromosomal integration if the target cell is postmitotic (e.g., muscle fibers) or slowly mitotic (e.g., hepatocytes) and if there is no immune response to the expressed gene (Wolff et al, 1992;Miao et al, 2000;Herweijer et al, 2001;Zhang et al, 2001;Wooddell et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Dose Response in Rodents and Nonhuman Primates After Hydrodynamic Limb Vein Delivery of Naked Plasmid DNA

et al. 2011

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The efficacy of gene therapy mediated by plasmid DNA (pDNA) depends on the selection of suitable vectors and doses. Using hydrodynamic limb vein (HLV) injection to deliver naked pDNA to skeletal muscles of the limbs, we evaluated key parameters that affect expression in muscle from genes encoded in pDNA. Short-term and long-term promoter comparisons demonstrated that kinetics of expression differed between cytomegalovirus (CMV), muscle creatine kinase, and desmin promoters, but all gave stable expression from 2 to 49 weeks after delivery to mouse muscle. Expression from the CMV promoter was highest. For mice, rats, and rhesus monkeys, the linear range for pDNA dose response could be defined by the mass of pDNA relative to the mass of target muscle. Correlation between pDNA dose and expression was linear between a threshold dose of 75 mg/g and maximal expression at approximately 400 mg/g. One HLV injection into rats of a dose of CMV-LacZ yielding maximal expression resulted in an average transfection of 28% of all hind leg muscle and 40% of the gastrocnemius and soleus. Despite an immune reaction to the reporter gene in monkeys, a single injection transfected an average of 10% of all myofibers in the targeted muscle of the arms and legs and an average of 15% of myofibers in the gastrocnemius and soleus.

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“…In this strategy, systemic administration has been intensively investigated assuming the liver, as a first-pass organ, exhibits strong abilities to retain DNA molecules; however, intraportal injection, direct injection of the gene therapy product in HCC has also been attempted. 69,70 Considering that the low gene transfer efficiency observed with this strategy would imply the injection of very large amounts of DNA molecules to efficiently hit HCC in vivo, numerous efforts are focused to improve delivery of the expression unit using chemical additives such as PEI, liposomes or polyamidoamine dendrimers, etc. in synergy or not with helper additives of viral origin.…”

Section: Nonviral Vectorsmentioning

confidence: 99%

Gene therapy of hepatocarcinoma: a long way from the concept to the therapeutical impact

Gerolami

Uch²,

Bréchot

et al. 2003

Cancer Gene Ther

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Time course of gene expression after plasmid DNA gene transfer to the liver

Abstract: The present data suggest that if promoter inactivation can be overcome, intravascular delivery of plasmid DNA could be a highly efficient, simple and non-toxic liver gene therapy approach. Intravascular delivery of pDNA allows for the rapid screening of novel expression vectors in vivo.

Cited by 132 publications

References 54 publications

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

Intranuclear disposition of exogenous DNA in vivo: Silencing, methylation and fragmentation

Dose Response in Rodents and Nonhuman Primates After Hydrodynamic Limb Vein Delivery of Naked Plasmid DNA

Gene therapy of hepatocarcinoma: a long way from the concept to the therapeutical impact

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