Exploiting viral cell-targeting abilities in a single polypeptide, non- infectious, recombinant vehicle for integrin-mediated DNA delivery and gene expression

Anna Arís, Jordi X. Feliu, Andrew Knight, Charles Coutelle, Antonio Villaverde

Research output: Contribution to journalArticleResearchpeer-review

31 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)689-696
JournalBiotechnology and Bioengineering
Volume68
Issue number6
DOIs
Publication statusPublished - 20 Jun 2000

Keywords

  • Cell targeting
  • FMDV
  • Integrin
  • Oligolysine
  • Recombinant protein
  • RGD

Fingerprint

Dive into the research topics of 'Exploiting viral cell-targeting abilities in a single polypeptide, non- infectious, recombinant vehicle for integrin-mediated DNA delivery and gene expression'. Together they form a unique fingerprint.

Cite this