Non-Viral Gene Delivery Vectors

Magdalena Wytrwal, Chantal Pichon

Cationic polymers have shown great potential for the delivery of proteins, nucleic acids forming complexes, called polyplexes. The most important issue in the context of using cationic polymers as carriers is the balance between the high transfection efficiency and low cytotoxicity. In this chapter, we report the preparation of polyallylamine derivatives mainly based on substitution of amino groups by glycidyltrimethylammonium chloride. The resulting polyplexes enhance the transfection of HeLa cell line without cytotoxic effects. Here, we describe methods for preparation and characterization of polyplexes using dynamic light scattering, ζ-potential measurements, gel retardation assay, and atomic force microscopy. Moreover, we provide protocols for the transfection of HeLa cell line by polyplexes, determination of their cytotoxicity, cell uptake, and intracellular trafficking.