Cell/gene-activated biomaterials are the main tendency and characteristics of the biomaterial researches for the future. The gene delivery based on the polymeric materials as vectors has potential applications in gene therapy, regenerative medicine and tissue engineering. In this paper, the application of layer-by-layer (LBL) technique in advanced biomaterial surface engineering is reviewed and its potential application in the construction of in situ gene delivery system onto biomaterial surface is in turn addressed. These are of great significance to biomaterial researches.