EFFICIENT TRANSFER OF GENETIC MATERIAL INTO MAMMALIAN-CELLS USING STARBURST POLYAMIDOAMINE DENDRIMERS

Starburst polyamidoamine dendrimers are a new class of synthetic polym ers with unique structural and physical characteristics. These polymer s were investigated for the ability to bind DNA and enhance DNA transf er and expression in a variety of mammalian cell lines. Twenty differe nt types of polyamidoamine dendrimers were synthesized, and the polyme r structure was confirmed using well-defined analytical techniques. Th e efficiency of plasmid DNA transfection using dendrimers was examined using two reporter gene systems: firefly luciferase and bacterial bet a-galactosidase. The transfections were performed using various dendri mers, and levels of expression of the reporter protein were determined . Highly efficient transfection of a broad range of eukaryotic cells a nd cell lines was achieved with minimal cytotoxicity using the DNA/den drimer complexes. However, the ability to transfect cells was restrict ed 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 enha nced transfection with the addition of chloroquine, indicating endosom al localization of the complexes. The capability of a dendrimer to tra nsfect cells appeared to depend on the size, shape, and number of prim ary amino groups on the surface of the polymer, However, the specific dendrimer most efficient in achieving transfection varied between diff erent types of cells. These studies demonstrate that Starburst dendrim ers can transfect a wide variety of cell types in vitro and offer an e fficient method for producing permanently transfected cell lines.