Enhanced Gene Delivery by Avidin-Displaying Baculovirus

6533b7d0fe1ef96bd125af25

RESEARCH PRODUCT

Enhanced Gene Delivery by Avidin-Displaying Baculovirus

Ari T. Marttila Varpu Marjomäki Olli H. Laitinen Kari J. Airenne Seppo Ylä-herttuala Markku S. Kulomaa Vesa P. Hytönen Anssi J. Mähönen Jani K. Raty Pauliina Lehtolainen

subject

Biotin binding Genetic Vectors Biotin Biosensing Techniques Biology Gene delivery Cell Line chemistry.chemical_compound Transduction (genetics)Biotin Viral envelope Transduction Genetic Cell Line Tumor Drug Discovery Genetics Animals Biotinylation Binding site Molecular Biology Pharmacology Epidermal Growth Factor Gene Transfer Techniques Avidin Molecular biology Cell biology Rats ErbB Receptors Spectrometry Fluorescence chemistry Biotinylation biology.protein Molecular Medicine Rabbits Baculoviridae Viral Fusion Proteins Avidin Protein Binding

description

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.

year	journal	country	edition	language
2004-02-01	Molecular Therapy

10.1016/j.ymthe.2003.11.004 http://dx.doi.org/10.1016/j.ymthe.2003.11.004