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Non-viral gene therapy vectors are highly desirable tools for the introduction of DNA into cells; they have better safety profiles than viral methods of delivery and are more amenable to repeated administration. Non-viral vectors include naked DNA, cationic lipid-DNA com‐ plexes (lipoplex), polymer-DNA complexes (polyplex) or combinations of lipids and polymers. Successful gene delivery depends upon the ability of the vector of choice to target a specific cell type, enter the cell and obtain sufficient levels of gene expression. This is not a simple task since there are several barriers encountered by both viral and non-viral vectors that make this process difficult. First, the vector must have a method by which to target a specific cell type, while also avoiding extracellular insults including nucleases and the immune system. Next, once the vector has reached its particular target, it must traverse the plasma membrane and/or escape the endosome, and pass through the dense cytoskeletal network en route to the nucleus. The nuclear envelope presents a final barrier, since DNA must enter the nucleus in order to be transcribed. While viruses have evolved mechanisms to enter target cells, deliver their genetic material and continue to propagate, non-viral systems lack these innate mecha‐ nisms. Consequently, there has been much work aimed at characterizing and overcoming these barriers in order to improve the efficacy of these vectors.

Extracellular and Intracellular Barriers to Non-Viral Gene Transfer