The aim of this work was to design a new non-viral carrier for gene delivery. This carrier was equipped with low molecular weight hyaluronic acid (LMHA) as ligand, which was conjugated by covalent attachment of hydrophobic amines (fatty amines) with different chain lengths and spermine as cationic segments. Their chemical structure and self-association behavior of the LMHA conjugates were investigated using 1H NMR, dynamic light scattering, fluorescence spectroscopy, and TEM. From the results, it was observed that polymeric micelles were spherical in shape. The micellar particle sizes and CMCs of the conjugates were significantly dependent on the degree of substitution of hydrophobic groups and the chain length. The CMCs of the conjugates were as low as 40–140 mg/L. The zeta potentials and agarose gel electrophoresis assays suggest that LMHA micelles encapsulating siRNA can be used as novel gene carriers for biomedical applications.