A recombinant, multifunctional protein has been designed for optimized, cell-targeted DNA delivery and gene expression in mammalian cells. This hybrid construct comprises a viral peptide ligand for integrin α(v)β3 binding, a DNA-condensing poly-L-lysine domain, and a complete, functional β-galactosidase protein that serves simultaneously as purification tag and DNA-shielding agent. This recombinant protein is stable; it has been produced successfully in Escherichia coli and can be purified in a single step by affinity chromatography. At optimal molar ratios, mixtures of this vector and a luciferase-reporter plasmid form stable complexes that transfect cultured cells. After exposure to these cell-targeted complexes, steady levels of gene expression are observed for more than 3 days after transfection, representing between 20 and 40% of those achieved with untargeted, lipid-based DNA- condensing agents. The principle to include vital motifs for cell infection in single polypeptide recombinant proteins represents a promising approach towards the design of non-viral modular DNA transfer vectors that conserve the cell-targeting specificity of native viruses and that do not need further processing after bioproduction in a recombinant host. (C) 2000 John Wiley and Sons, Inc.
|Journal||Biotechnology and Bioengineering|
|Publication status||Published - 20 Jun 2000|
- Cell targeting
- Recombinant protein