Delivering genes into cells or tissues is a very promising tool in treatment of genetic diseases and cancer. In the area of gene therapy, the investigation is focused on the development of efficient viral and nonviral transfection vectors. Among nonviral vectors, cationic liposome mediated transfection holds considerable promise for its (a) reproducibility and simplicity in preparation, (b) nonimmunogenic nature, and (c) efficiency in forming stable complexes even with large DNA. A number of lipids were synthesized and their transfection efficiencies were tested with success. The key structural elements of these transfection lipids include the presence of a hydrophobic group either directly linked to an amino headgroup or connected by a spacer molecule to a positively charged amino acid via an amide bond. The hydrophobic molecule parts were synthesized starting from four different compounds: malonic acid ester, glycerol, pentaerythritol and tris(hydroxymethyl)aminomethan. In general, these molecules were selectively alkylated with long chain alkylhalides using a blocking group strategy. On the basis of the starting material there are one or two free functional sites left after the alkylation process. The polyamine building blocks, the spacer or an amino acid were coupled to these free sites using peptide-coupling chemistry. The synthetic scheme can easily be extended to give lipids of different charge, spacer or lipid chain length. The synthesized lipids were characterized by monolayer investigation including film balance measurements. Finally transfection efficiency and cellular damage were characterised, size and stability of the liposomes were determined. It could be shown, that the lipids are particularly suitable for gene transfection with respect to simplicity of preparation, efficiency and lack of specific immune response.