Targetable Endolytic Protein-Based Polymers for Systemic Breast Cancer Gene Therapy

摘要

Using recombinant techniques the first prototype polymer-protein fusion was biosynthesized to mediate transgene expression in mammalian cell lines. The carrier was designed to have the structure of (KHKHKHKHKK)6-FGF2 where Lysine (K) residues would allow complexation with plasmid DNA by electrostatic interactions, basic Fibroblast Growth Factor (FGF2) to target cells over-expressing FGF2 receptors (FGFR), and Histidine (H) residues to facilitate escape from the endosomoal compartments due to proton sponge effect. The gene carrier was biosynthesized in E.coli, purified, characterized, complexed with pDNA (pEGFP) and the complexes were used to transfect mammalian cells. Results demonstrate the successful cloning, expression, purification and bioactivity of the carrier. The carrier/DNA complexes transfected mammalian cells known to express FGFR. Genetic engineering techniques show promise for systematic investigation of structure-activity relationship of non-viral polymeric vectors.