Exploiting viral cell-targeting abilities in a single polypeptide, non-infectious, recombinant vehicle for integrin-mediated DNA delivery and gene expression
✍ Scribed by Anna Arís; Jordi X. Feliu; Andrew Knight; Charles Coutelle; Antonio Villaverde
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 201 KB
- Volume
- 68
- Category
- Article
- ISSN
- 0006-3592
No coin nor oath required. For personal study only.
✦ Synopsis
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 ␣ V  3 binding, a DNA-condensing poly-L-lysine domain, and a complete, functional -galactosidase protein that serves simultaneously as purification tag and DNAshielding 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 celltargeted 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 viral 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.