Dramatic effect of Mg2+ on transfecting mammalian cells by DNA/calcium phosphate precipitates

Chowdhury, Ezharul Hoque; Kunou, Megumi; Harada, Ichiro; Kundu, Anup K.; Akaike, Toshihiro

doi:10.1016/j.ab.2004.01.009

Cited by 15 publications

(16 citation statements)

References 9 publications

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“…The estimated molar ratios of Ca, Mg and P present in the precipitates of Ca/MgPi indicated formation hydroxyapatite with the molecular formula of Ca 10−X Mg X (PO 4 ) 6 (OH) 2 for 0.58 and 1.03 percentage of Mg and octacalcium phosphate (OCP) with the formula of Ca 4−X Mg X (PO 4 ) 3 for 1.76 to 3.16 percentage of Mg substituted in the particles [13]. The uptake of fluorescence-labeled plasmid in HeLa cells indicated size-dependent endocytosis of the Ca/MgPi particles with the particles of the smaller size resulting in higher uptake of the DNA compared to those of bigger size, implying in agreement with the notion that internalization of apatite particles is indeed size-dependent [18,19].…”

Section: +supporting

confidence: 74%

pH-responsive magnesium- and carbonate-substituted apatite nano-crystals for efficient and cell-targeted delivery of transgenes

Chowdhury¹

2013

OJGen

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The short half-lives due to the enzymatic degradation in blood, the lack of tissue targetability and the incapability to passively diffuse across the plasma membrane and smoothly traffic across the harsh intracelluar environment are the major shortcomings for nucleic acid-based potential therapeutics, such as recombinant plasmid and antisense oligonucleotides or small interferring RNA (siRNA). Plasmid DNA containing a gene of interest could have immense impact as a promising therapeutic drug for treating genetic as well as acquired human diseases at the molecular level with high level of efficacy and precision. Thus both viral and non-viral synthetic vectors have been developed in the past decades to address the aforementioned challenges of naked DNA. While in the viral particles plasmid DNA is integrated into the viral genome, in most non-viral cases the DNA being anionic in nature is electrostatically associated with a cationic lipid or polymer forming lipoplex or polyplex, respectively, or a cationized inorganic gold, silica or iron oxide particle. Due to the potential immunogenicity and carcinogenicity issues with the viral particles, non-viral vectors have drawn much more attention for the clinical evaluation. However, the main concern of using non-biodegradable particles, specially the inorganic ones, is the adverse effects owing to their long term interactions with body components. We have recently developed biodegradable pH-sensitive inorganic nanoparticles of Mg/CaPi and carbonate apatite for efficient transgene delivery to primary, cancer and embryonic stem cells, by virtue of their high affinity binding with the DNA, ability to contact the cell membrane by ionic or ligand-receptor interactions and fast dissolution kinectis in endosomal acidic pH facilitating release of the DNA from the dissolving particles and also from the endosomes.

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Section: +supporting

confidence: 74%

pH-responsive magnesium- and carbonate-substituted apatite nano-crystals for efficient and cell-targeted delivery of transgenes

Chowdhury¹

2013

OJGen

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“…The biological activity of DNA was retained even after the ACP-assisted biomimetic coating process (see figures [4][5][6], as reported previously [15][16][17][18]. This retention of activity is due to the mild coating conditions: neutral pH, normal pressure and temperature, and enzyme-free acellular environment.…”

Section: Discussionsupporting

confidence: 48%

“…It has emerged as an important tool even in tissue engineering applications. Among conventional gene transfer systems, a calcium phosphate system [1][2][3][4][5][6] that is mediated by nano-and microscale particles of DNA-apatite composites is superior in safety and biocompatibility to other alternatives, such as viral [7][8][9] and lipid [10][11][12] systems. However, the gene transfer efficiency of the calcium phosphate system is relatively low.…”

Section: Introductionmentioning

confidence: 99%

Fabrication of DNA-antibody–apatite composite layers for cell-targeted gene transfer

Yazaki

Oyane

Araki

et al. 2012

Science and Technology of Advanced Materials

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Surface-mediated gene transfer systems using apatite (Ap)-based composite layers have received increased attention in tissue engineering applications owing to their safety, biocompatibility and relatively high efficiency. In this study, DNA-antibody-apatite composite layers (DA-Ap layers), in which DNA and antibody molecules are immobilized within a matrix of apatite nanocrystals, were fabricated using a biomimetic coating process. They were then assayed for their gene transfer capability for application in a specific cell-targeted gene transfer. A DA-Ap layer that was fabricated with an anti-CD49f antibody showed a higher gene transfer capability to the CD49f-positive CHO-K1 cells than a DNA-apatite composite layer (D-Ap layer). The antibody facilitated the gene transfer capability of the DA-Ap layer only to the specific cells that were expressing corresponding antigens. When the DA-Ap layer was fabricated with an anti-N-cadherin antibody, a higher gene transfer capability compared with the D-Ap layer was found in the N-cadherin-positive P19CL6 cells, but not in the N-cadherin-negative UV♀2 cells or in the P19CL6 cells that were pre-blocked with anti-N-cadherin. Therefore, the antigen-antibody binding that takes place at the cell-layer interface should be responsible for the higher gene transfer capability of the DA-Ap than D-Ap layer. These results suggest that the DA-Ap layer works as a mediator in a specific cell-targeted gene transfer system.

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“…Conventional gene transfer systems are generally mediated by particulate transfection reagents such as viruses [1][2][3], liposomes [4][5][6], and calcium phosphates [7][8][9][10]. These particle-mediated gene transfer systems are, however, generally weak in spatial control of gene transfer.…”

Section: Introductionmentioning

confidence: 99%

Fabrication of a DNA-lipid-apatite composite layer for efficient and area-specific gene transfer

Oyane

Yazaki

Araki

et al. 2012

J Mater Sci: Mater Med

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A surface-mediated gene transfer system using biocompatible apatite-based composite layers has great potential for tissue engineering. Among the apatite-based composite layers developed to date, we focused on a DNA-lipid-apatite composite layer (DLp-Ap layer), which has the advantage of relatively high efficiency as a non-viral system. In this study, various lipid transfection reagents, including a newly developed reagent, polyamidoamine dendron-bearing lipid (PD), were employed to prepare the DLp-Ap layer, and the preparation condition was optimized in terms of efficiency of gene transfer to epithelial-like CHO-K1 cells in the presence of serum. The optimized DLp-Ap layer derived from PD had the highest gene transfer efficiency among all the apatite-based composite layers prepared in this study. In addition, the optimized DLp-Ap layer demonstrated higher gene transfer efficiency in the presence of serum than the conventional particle-mediated systems using commercially available lipid transfection reagents. It was also shown that the optimized DLp-Ap layer mediated the area-specific gene transfer on its surface, i.e., DNA was preferentially transferred to the cells adhering to the surface of the layer. The present gene transfer system using the PD-derived DLp-Ap layer, with the advantages of high efficiency in the presence of serum and area-specificity, would be useful in tissue engineering.

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Dramatic effect of Mg2+ on transfecting mammalian cells by DNA/calcium phosphate precipitates

Cited by 15 publications

References 9 publications

pH-responsive magnesium- and carbonate-substituted apatite nano-crystals for efficient and cell-targeted delivery of transgenes

pH-responsive magnesium- and carbonate-substituted apatite nano-crystals for efficient and cell-targeted delivery of transgenes

Fabrication of DNA-antibody–apatite composite layers for cell-targeted gene transfer

Fabrication of a DNA-lipid-apatite composite layer for efficient and area-specific gene transfer

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