Non-viral neuronal gene delivery mediated by the HC fragment of tetanus toxin

Many inherited neurological diseases and cancers could potentially benefit from efficient targeted gene delivery to neurons of the central nervous system. The nontoxic fragment C (H-C) of tetanus toxin retains the specific nerve cell binding and transport properties of tetanus holotoxin. The H-C fragment has previously been used to promote the uptake of attached proteins such as horseradish peroxidase, beta-galactosidase and superoxide dismutase into neuronal cells in vitro and in vivo. We report the use of purified recombinant H-C fragment produced in yeast and covalently bound to polylysine [poly(K)] to enable binding of DNA. We demonstrate that when used to transfect cells, this construct results in nonviral gene delivery and marker gene expression in vitro in N18 RE 105 cells (a neuroblastoma x glioma mouse/rat hybrid cell line) and F98 (a glioma cell line). Transfection was dependent on H-C and was neuronal cell type specific. H-C may prove a useful targeting ligand for future neuronal gene therapy.