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J. Biol. Chem., Vol. 266, Issue 22, 14338-14342, 08, 1991
GY Wu, JM Wilson, F Shalaby, M Grossman, DA Shafritz and CH Wu
A plasmid (palb3) was constructed containing the structural gene for human
serum albumin driven by mouse albumin enhancer-rat albumin promoter
elements. Using an asialoglycoprotein-polycation conjugate consisting of
asialoorosomucoid coupled to poly-L-lysine, a soluble DNA complex was
formed that was capable of targeting specifically to hepatocytes via
asialoglycoprotein receptors present on these cells. Groups of Nagase
analbuminemic rats were injected with complexed DNA or controls, followed
by two-thirds partial hepatectomy to stimulate hepatocyte replication.
Using a cDNA probe for the human albumin structural gene, hybridizable
sequences were detected in analbuminemic rats treated with complex as
determined by Southern blot analysis. Two weeks post-injection, the
targeted DNA was found to exist primarily in plasmid form with an average
copy number of 1000/diploid cell. Human albumin mRNA was detected by
dot-blot hybridization with a specific oligonucleotide cDNA probe and
confirmed by RNase protection assay using a vector-specific probe.
Circulating human albumin was detected in the serum of palb3-treated Nagase
analbuminemic rats by Western blots using an antibody specific for human
serum albumin. A time course demonstrated that circulating human albumin
was not detectable 24 h after injection, but became measurable at a level
of 0.05 micrograms/ml within 48 h and increased in concentration to a
maximum of 34 micrograms/ml by 2 weeks post-injection. This level of
expression remained stable through 4 weeks after injection and partial
hepatectomy.
Receptor-mediated gene delivery in vivo. Partial correction of genetic analbuminemia in Nagase rats
Department of Medicine, University of Connecticut School of Medicine, Farmington 06030.
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