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After revisiting the captioned problem by using a combination of chemical synthesis and physical methods, we studied the dynamics of the complexation between branched polyethyleneimine (bPEI) and plasmid DNA (pDNA) and characterized the structure, size and surface charge of the resultant DNA/PEI complexes (polyplexes). As expected, in order to reach a high efficiency in the gene transfection into cells, it is necessary to use a higher N ratio and make the polyplexes positively charged. Our results reveal that it is those uncomplexed bPEI chains free in the solution mixture that play a vitally important role in promoting the gene transfection, leading to some new thinking of how to correlate in vitro and in vivo studies so that we will be able to improve the in vivo transfection efficiency. Increasing the N ratio normally results in a higher cytotoxicity, which is a catch-22 problem. Recently, we found that a proper modification of bPEI can greatly reduce its cytotoxicity without any suffering in the transfection efficiency. In this lecture, we will show that our properly modified bPEI is even much more effective and less cytotoxic in the in vitro gene transfection than those commercially available lipoplexes. Our recent new development leads to a completely new direction in the development of non-viral vectors for molecular medicines, including the gene transfection.
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