Efficient transfer of genetic material into mammalian cells using Starburst polyamidoamine dendrimers.

Abstract

Starburst polyamidoamine dendrimers are a new class of synthetic polymers with unique structural and physical characteristics. These polymers were investigated for the ability to bind DNA and enhance DNA transfer and expression in a variety of mammalian cell lines. Twenty different types of polyamidoamine dendrimers were synthesized, and the polymer structure was confirmed using welldefined analytical techniques. The efficiency of plasmid DNA transfection using dendrimers was examined using two reporter gene systems: firefly luciferase and bacterial 1B-galactosidase. The transfections were performed using various dendrimers, and levels of expression of the reporter protein were determined. Highly efficient transfection of a broad range of eukaryotic cells and cell lines was achieved with minimal cytotoxicity using the DNA/dendrimer complexes.However, the ability to transfect cells was restricted to certain types of dendrimers and in some situations required the presence of, additional compounds, such as DEAE-dextran, that appeared to alter the nature of the complex. A few cell lines demonstrated enhanced transfection with the addition of chloroquine, indicating endosomal localization of the complexes. The capability of a dendrimer to transfect cells appeared to depend on the size, shape, and number of primary amino groups on the surface of the polymer. However, the specific dendrimer most efficient in achieving transfection varied between different types of cells. These studies demonstrate that Starburst dendrimers can transfect a wide variety of cell types in vitro and offer an efficient method for producing permanently transfected cell lines.

Polyphosphoramidate gene carriers: effect of charge group on gene transfer efficiency

“…In contrast, the enhancement effect on the other three carriers was not obvious (Figure 8b). In addition, the enhancement effect of CQ on PPA-EA/DNA nanoparticle transfection was only pronounced at lower charge ratios (6)(7)(8)(9)(10)(11)(12). No enhancement was observed at charge ratio above 15.…”

Highly efficient suicide gene expression in hepatocellular carcinoma cells by Epstein-Barr virus-based plasmid vectors combined with polyamidoamine dendrimer

“…27 In our first series of experiments, several different ratios of DNA to PAAD were tested for HLE cells (Fig 3A). The highest ␤-gal activity was observed when 15 g of PAAD and 2 g of pSES.␤ were used.…”

Co-polymer of histidine and lysine markedly enhances transfection efficiency of liposomes

“…[27][28][29][30] These include polyethylenimine, Superfect, and polyamidoamine whose single repeating subunit contains both a positive charge and buffering capacity. Since binding and buffering properties within these polymers are fixed and cannot be varied, we believe that there is an inherent inflexibility of a polymer that has a single repeating subunit with two or more functional features.…”