Dominating issues in gene vector optimization are specific in recognizing the target cells and exploiting the proper intracellular trafficking routes. Any progress in this area will result in improved specific gene transfer, reduce the required therapeutic vector doses and, in consequence, lower the overall toxicity to the host. To provide polyplexes with the ability to distinguish between non-target and target cells, cell-binding ligands have been incorporated which recognize target-specific cellular receptors. In addition, polyplex domains with unspecific binding capacity (such as surface charges) have to be shielded or removed. Cell-binding ligands can be small molecules, vitamins, carbohydrates, peptides or proteins such as growth factors or antibodies. Such ligands have been incorporated into polyplexes after chemical conjugation to cationic polymers. The choice of the ligand and physical properties of the DNA formulation strongly influence extracellular routing (circulation in blood, tissue distribution), uptake and intracellular delivery of polyplexes. Recent efforts are discussed that aim at the development of polyplexes into virus-like supramolecular complexes; such particles should undergo structural changes compatible with extracellular and intracellular targeting.