Magnetic lamellar hydroxyapatite (ML-HA) nanoparticles were synthesized by a template-assisted self-assembly process. The as-prepared ML-HA nanoparticles self-assembled under different conditions were characterized by XRD, TEM, cytotoxicity assessment, and DNA-loading and transfection efficiency measurements. We found that the structure and morphology of ML-HA were controlled by self-assembly conditions. The ML-HA synthesized in this work exhibited good biocompatibility. The DNA-loading capacity and zeta-potential of ML-HA were much lower in comparison to bare lamellar HA (L-HA) without magnetic nanoparticles. Despite that, the ML-HA with good lamellar structure showed 47% higher transfection efficiency than L-HA. Results suggested that the ordered lamellar structure is a key factor in controlling transfection efficiency and magnetization is an effective way of improving the transfection efficiency of lamellar HA. Mechanisms were proposed to interpret these experimental results. It is demonstrated that the ML-HA may be a promising gene vector to deliver DNA into the cells effectively and safely. (C) 2015 Elsevier B.V. All rights reserved.