Enhanced gene delivery by avidin-displaying baculovirus

Mol Ther. 2004 Feb;9(2):282-91. doi: 10.1016/j.ymthe.2003.11.004.

Abstract

Flexible alteration of virus surface properties would be beneficial for enhanced and targeted gene delivery. A useful approach could be based on a high-affinity receptor-ligand pair, such as avidin and biotin. In this study, we have constructed an avidin-displaying baculovirus, Baavi. Avidin display was expected to enhance cell transduction due to the high positive charge of avidin in physiological pH and to provide a binding site for covering the virus with desired biotinylated ligands. Successful incorporation of avidin on the virus envelope was detected by immunoblotting and electron microscopy. Multiple biotin-binding sites per virus were detected with fluorescence-correlation spectroscopy and tight biotin binding was observed using an optical biosensor, IAsys. Baavi showed a 5-fold increase in transduction efficiency in rat malignant glioma cells (BT4C) and a 26-fold increase in rabbit aortic smooth muscle (RAASMC) cells compared to wild-type baculovirus. Enhanced transduction was also observed with biotinylated target cells. Biotinylated epidermal growth factor (EGF) enabled specific targeting of the virus with high efficiency to EGF receptor-expressing (SKOV-3) cells. An additional advantage of the avidin display was demonstrated with biotinylated paramagnetic particles, which enabled magnetic targeting. Altogether, we show that avidin display is a rapid and versatile method to improve viral properties for gene delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Avidin / genetics
  • Avidin / metabolism*
  • Baculoviridae / genetics*
  • Baculoviridae / physiology*
  • Biosensing Techniques
  • Biotin / metabolism
  • Biotinylation
  • Cell Line
  • Cell Line, Tumor
  • Epidermal Growth Factor / metabolism
  • ErbB Receptors / metabolism
  • Gene Transfer Techniques*
  • Genetic Vectors / genetics
  • Genetic Vectors / physiology
  • Protein Binding
  • Rabbits
  • Rats
  • Spectrometry, Fluorescence
  • Transduction, Genetic
  • Viral Fusion Proteins / genetics
  • Viral Fusion Proteins / metabolism

Substances

  • Viral Fusion Proteins
  • biotinyl epidermal growth factor
  • Avidin
  • Epidermal Growth Factor
  • Biotin
  • ErbB Receptors