Nuclear‐targeted minicircle to enhance gene transfer with non‐viral vectors <i>in vitro</i> and <i>in vivo</i>

Vaysse, Laurence; Gregory, Lisa G.; Harbottle, Richard P.; Pérouzel, Éric; Tolmachov, Oleg; Coutelle, Charles

doi:10.1002/jgm.883

Cited by 45 publications

(33 citation statements)

References 38 publications

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“…[33] One type of cationic lipid that is often used in DNA transfection is DOTAP. [34][35][36] Using this molecule, we have elaborated a clever strategy for the production of cationic MLs, which was outlined in a previous paper.…”

Section: Discussionmentioning

confidence: 99%

Optimal Conditions for Labelling of 3T3 Fibroblasts with Magnetoliposomes without Affecting Cellular Viability

2007

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A comparative study that deals with the internalisation of different types of magnetoliposomes (MLs) by 3T3 fibroblasts revealed that cationic MLs proved to be superior to neutral and anionic ones. Internalisation was visualised both by optical light and transmission electron microscopy. The latter showed that the cationic MLs ultimately ended up in lysosomal structures. The effect of increasing 1,2-dioleoyl-3-trimethylammonium propane (DOTAP) concentrations in the cationic ML coat has been elucidated. High uptake efficiency was only achieved with MLs that carry a high DOTAP payload. However, these structures also demonstrated toxic effects. The use of the saturated distearoyl analogue (DSTAP) at identical concentrations led to improved uptake efficiency and lower toxicity. By using iron-oxide-free vesicles, it was shown that the toxicity was due to lipid bilayer constituents and not the iron oxide. In conclusion, the use of DMPC-DSTAP (96.67:3.33; molar ratio) MLs results in an extremely high labelling of 3T3 fibroblasts with iron oxides (47.66 pg Fe per cell) without evoking any influence on cell viability.

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Section: Discussionmentioning

confidence: 99%

Optimal Conditions for Labelling of 3T3 Fibroblasts with Magnetoliposomes without Affecting Cellular Viability

2007

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“…Recently, one study analyzed the ability of a tetracycline repressor protein TetR fused to the TAT peptide to enhance nuclear import and gene expression of minicircle DNA in vivo. It was found that there was more than a 6-fold increase in gene expression in mouse lung [Vaysse et al 2006]. Another HIV protein, the integrase IN, has shown nuclear accumulation and DNA binding activity in vitro, and may be a candidate for enhancement of nonviral gene delivery [Hearps & Jans 2006].…”

Section: Nls:dna Conjugatesmentioning

confidence: 99%

“…When EBV nuclear antigen (EBNA)-1 expressing cells were cytoplasmically microinjected with plasmids containing the oriP DTS, increased gene expression was detected when compared to plasmids lacking oriP [Langle-Rouault et al 1998]. More recently, combinations of the tet operator and a modified tetracycline repressor containing an NLS (tetO and TetR-NLS, respectively) have been used in cis and trans to show that nuclear import can be controlled and enhanced by protein-DNA interactions [Vaysse et al 2004;Vaysse et al 2006]. When multiple copies of the tetO sequence were cloned into a plasmid and transfected into cells expressing TetR-NLS, gene expression increased almost 20-fold in growth-arrested cells, and nuclear localization increased by 4-fold.…”

Section: Sequence-specific Nuclear Import Of Plasmid Dnamentioning

confidence: 99%

Intracellular Trafficking of Plasmids for Gene Therapy: Mechanisms of Cytoplasmic Movement and Nuclear Import

Vaughan¹,

DeGiulio²,

Dean³

2006

CGT

107

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Under physiologically relevant conditions, the levels of non-viral gene transfer are low at best. The reason for this is that many barriers exist for the efficient transfer of genes to cells, even before any gene expression can occur. While many transfection strategies focus on DNA condensation and overcoming the plasma membrane, events associated with the intracellular trafficking of the DNA complexes have not been as extensively studied. Once internalized, plasmids must travel potentially long distances through the cytoplasm to reach their next barrier, the nuclear envelope. This review summarizes the current progress on the cytoplasmic trafficking and nuclear transport of plasmids used for gene therapy applications. Both of these processes utilize specific and defined mechanisms to facilitate movement of DNA complexes through the cell. The continued elucidation and exploitation of these mechanisms will lead to improved strategies for transfection and successful gene therapy.

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“…To address this problem, one important issue is how to overcome the inefficient translocation of plasmid entities to cell nucleus. It is possible to attach the plasmid cargos with a nuclear localization signal peptide to conquer the barrier (Vaysse et al, 2006). Finally, the presence of bacterial DNAs naturally rich in CpG dinucleotides on plasmids is known to cause transcriptional silencing of the transgene in cells.…”

Section: Discussionmentioning

confidence: 99%

Engineering of Escherichia coli for targeted delivery of transgenes to HER2/neu‐positive tumor cells

Chang

Cheng

Chen

et al. 2011

Biotech & Bioengineering

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Targeting of non-phagocytic tumor cells and prompt release of gene cargos upon entry into tumors are two limiting steps in the bacterial gene delivery path. To tackle these problems, the non-pathogenic Escherichia coli strain BL21(DE3) was engineered to display the anti-HER2/neu affibody on the surface. After co-incubation with tumor cells for 3 h, the anti-HER2/neu affibody-presenting E. coli strain was selectively internalized into HER2/neu-positive SKBR-3 cells. The invasion efficiency reached as high as 30%. Furthermore, the bacteria were equipped with the phage ϕX174 lysin gene E-mediated autolysis system. Carrying the transgene (e.g., eukaryotic green fluorescent protein, GFP), the tumor-targeting bacteria were subjected to the thermal shock to trigger the autolysis system upon entry into HER2/neu-positive cells. Flow cytometric analysis revealed that 3% of infected cells expressed GFP 24 h post thermal induction. Overall, the results show a promise of the proposed approach for developing bacteria as a delivery carrier.

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Nuclear‐targeted minicircle to enhance gene transfer with non‐viral vectors in vitro and in vivo

Abstract: Our results indicate that the combination of a minicircle DNA construct with a TetR nuclear-targeting system is able to potentiate gene expression of non-viral vectors.

Cited by 45 publications

References 38 publications

Optimal Conditions for Labelling of 3T3 Fibroblasts with Magnetoliposomes without Affecting Cellular Viability

Optimal Conditions for Labelling of 3T3 Fibroblasts with Magnetoliposomes without Affecting Cellular Viability

Intracellular Trafficking of Plasmids for Gene Therapy: Mechanisms of Cytoplasmic Movement and Nuclear Import

Engineering of Escherichia coli for targeted delivery of transgenes to HER2/neu‐positive tumor cells

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