Two novel micelle-forming amino-functionalized lipids (OT6 and TT6) bearing two alkyl chains connected to a large positively charged hexavalent headgroup, which might be interesting polynucleotide transferring agents with the advantage of an easy and reproducible production of micelle dispersions, have been characterized. The critical micelle concentration (cmc) of both lipids has been determined by two different methods, namely, isothermal titration calorimetry (ITC) and 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence experiments. In addition, the lipid dispersions were studied as a function of temperature using differential scanning calorimetry (DSC), dynamic light scattering (DLS), Fourier-transform infrared (FT-IR) spectroscopy, and cryo-transmission electron microscopy (cryo-TEM). The OT6 and TT6 micelles effectively complex DNA as determined by ITC and DSC measurements. In addition, DLS and ζ-potential measurements were performed to determine lipoplex formulations that exhibit colloidal stability. Finally, the structures of OT6/DNA complexes were investigated by means of X-ray scattering and TEM.