Specific and efficient targeting of adenovirus vectors to macrophages: Application of a fusion protein between an adenovirus-binding fragment and avidin, linked to a biotinylated oligonucleotide
article
Background: The application of serotype 5 adenoviruses (Ad5) in macrophages is hampered by the absence of the endogenous coxsackie adenovirus receptor (CAR). Methods: To overcome this limitation, we first generated a linker protein consisting of the virus-binding-domain of CAR and the C-terminus of avidin. Second, to target macrophages, this linker protein was equipped with the biotinylated (bio) oligonucleotide dA6G10, which was previously shown to display a high affinity for the scavenger receptor A (SR-A). Results: As compared to nontargeted virus, the linke r protein equipped with bio-dA6G10 showed a 500-fold increased reporter gene expression in mouse macrophage RAW264.7 cells. A linker protein equipped with a bio-dA16 control oligonucleotide was inactive. Moreover, the bio-dA6G10-equipped linker showed a 390-fold increased luciferase expression in the macrophage cell line J774 and 276- and 150-fold increased reporter gene expression in primary peritoneal and bone marrow (BM)-derived macrophages, respectively. Using BM-derived macrophages from SR-A knockout mice, it was shown that the dA6G10-mediated uptake is predominantly SR-A-mediated. Conclusions: Thus, we have developed a novel tool to link biotinylated ligands to a virus-binding fragment of CAR and have exploited this linker protein to extend the applicability of Ad5 to infect transformed and primary macrophages. Copyright © 2006 John Wiley & Sons, Ltd. Chemicals / CAS: Avidin, 1405-69-2; CAR receptor; Ligands; Oligonucleotides; Receptors, Virus; Recombinant Fusion Proteins
Topics
AtherosclerosisGene therapyMacrophagesTargetingadenovirus vectoravidincoxsackie virus and adenovirus receptorhybrid proteinluciferaseoligonucleotidescavenger receptoranimal cellanimal experimentbinding affinitybiotinylationbone marrow cellcarboxy terminal sequencecontrolled studyCoxsackie virusculture mediumgene targetinggenetic transfectionhuman cellin vivo studymacrophagemolecular cloningmousenonhumanperitoneum cellprotein analysisprotein bindingprotein domainprotein expressionprotein purificationprotein synthesisreporter geneserotypeviral gene delivery systemAdenoviridaeAnimalsAvidinBiotinylationCercopithecus aethiopsCOS CellsFemaleGene TargetingGene TherapyGenetic VectorsHela CellsHumansLigandsMacrophagesMiceOligonucleotidesReceptors, VirusRecombinant Fusion Proteins
TNO Identifier
239292
ISSN
1099498X
Source
Journal of Gene Medicine, 8(6), pp. 668-678.
Pages
668-678
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