C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/87 (2006.01)
Patent
CA 2090355
The present invention relates to the field of introducing genetic material to cultured cells and into cells of animals in vivo. In particular, the invention relates to the use of receptor-targeted neoglycoprotein to carry genetic material into cells. The invention also uses nuclear-localization signals(Gerace et al. Annu. Rev. Cell Biol. 1988, 4:335-374), to facilitate the entry of genetic materials into cell nucleus for gene expression. The present invention has a wide variety of applications, for example, in treatment of genetic diseases, in study of cell biology, and in genetic interventions of physiological and pathological processes. The present invention is a effective method for introducing and expressing genes in hepatoma cells and hepatocytes in vivo. Its design has several novel features. First, this invention uses a simple coupling reagent (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) to chemically add galactose to a protein, histone, in a one step reaction. The galactosylated protein (for example, albumin, histone and any other protein) has high affinity for the ASGP receptor; Second, the invention uses a protein which is a natural DNA-binding protein; Three, this invention uses a protein which crosses the membranes of cell nucleus by a facilitated process in addition to simple diffusion. This feature contributes greatly to the high level expression of the introduced foreign genes; Last, the galactosylated histone 1 is the physical embodiment of all the features in this invention. This means this gene delivery system has only one component and is simple, reliable, and efficient from engineering-design point of view.
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