Synthesis of a non-viral vector for gene transfer via the high-affinity neurotensin receptor.
We describe herein a method for synthesizing a non-viral gene vector that exploits the internalization properties of neurotensin (NT), as well as the procedures for a successful gene transfer to cells via the high-affinity NT receptor. The gene vector is NT cross-linked with poly-L-lysine via N-succinimidyl-6-[3'-(2-pyridyldithio)propionamido]hexanoate (LC-SPDP). The SPDP-derivatives containing either NT or poly-L-lysine are purified by gel filtration. The non-viral vector resulting from the reaction of NT-SPDP with HS-SPDP-poly-L-lysine is purified on Biogel A-1.5 m. This vector is complexed with plasmid DNA at a specific molar ratio to form the NT-polyplex, which ensures the delivery of the gene of interest to cells under conditions of receptor-mediated internalization. The NT-polyplex has shown ability to mediate transient gene expression in vitro [Brain Res. Mol. Brain Res. 69 (1999) 249] and in vivo [Soc. Neurosci. Abstr. 25 (1999) 67. 7]. This approach holds great promise for research and therapy.
Martinez-Fong
D
D
Departamento de Fisiología, Biofísica y Neurociencias; Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional de México, Apartado postal 14-740, 07000 D.F., México, Mexico. dmartine@fisio.civestav.mx
Navarro-Quiroga
I
I
D001667:Binding, Competitive; D003432:Cross-Linking Reagents; D004247:DNA / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000138:chemical synthesis*; D011107:Polylysine; D018028:Receptors, Neurotensin / Q000378:metabolism*; D013388:Succinimides; D014407:Tumor Cells, Cultured
D003432:Cross-Linking Reagents; D018028:Receptors, Neurotensin; D013388:Succinimides; D011107:Polylysine; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D004247:DNA