High-Efficiency Gene Transfer Mediated by Adenovirus Coupled to DNA–Polylysine Complexes

Abstract

Employment of recombinant viruses as gene transfer vectors is limited by constraints on the size and functional design of the genetic material to be transferred as well as potential safety hazards deriving from obligatory co-transfer of viral genetic elements. As an alternative strategy that capitalizes on the efficient cellular entry mechanisms of viruses, we have derived adenovirus-polylysine-DNA complexes whereby foreign DNA is transferred bound to the exterior of the virion. This linkage was accomplished utilizing an antibody bridge in which a monoclonal antibody was rendered competent to carry DNA by the attachment of a polylysine residue. Attachment of the antibody-polylysine to the virus was by virtue of the antibody's specificity for the virion. The resulting vector system mediates high-efficiency gene transfer to target cells in vitro. In addition, this vector design allows greatly enhanced flexibility in terms of the size and design of heterologous sequences that can be transferred. Since this strategy selectively exploits viral entry functions, which are independent of viral gene expression, the potential exists to derive vectors that avoid the hazards deriving from transfer of parent virus genome.

Antineoplastic effect of anti-erbB-2 intrabody is not correlated with scFv affinity for its target

“…To this end, we have explored the utility of efficient gene delivery vehicles in the context of intrabody knockout strategies. [27][28][29][30] In addition, our group and others have confirmed the key importance of using heterologous cellular processing signals to achieve effective intrabody colocalization with target oncoproteins. 16 -24,30 -32 In the present study, we sought to determine whether improvement in the affinity of the parent scFv could offer an additional means to improve the efficiency of intrabody-mediated oncoprotein knockout.…”

Modifications of the fiber in adenovirus vectors increase tropism for malignant glioma models

“…Polylysine has been shown to interact with heparan sulfate as well as other polyanions such as DNA, chondroitin sulfate, and mucins. 18,19 Because these molecules are expressed on a variety of cell types, AdZ.F(pK7) acts as a nonspecific vector that can transduce a broad spectrum of tissues. 9 Because different glioma cell lines have been shown to express these ubiquitous cellular receptors, AdZ.F(pK7) may transduce an expanded range of glioma cell lines at higher efficiencies than do unmodified vectors.…”

Enhancement of calcium phosphate-mediated transfection by inclusion of adenovirus in coprecipitates

“…As indicated above, adenovirus has been incorporated into other gene transfer vectors, including cationic lipids and receptortargeted polylysine complexes. [5][6][7][8]10,12,14 In these cases adenovirus is also thought to facilitate escape of the plasmid from the endosome.…”

“…A number of methods have been developed to facilitate plasmid-mediated gene transfer, including direct injection, electroporation, and incorporation of plasmid in complexes with cationic lipids, polylysine, and other molecules. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15] The use of calcium phosphate (CaPi) to coprecipitate DNA was one of the first methods used for DNA transfer to cells. 16 CaPi, as well as cationic lipids and other cationic polymers, condense the DNA and facilitate its uptake into the cell.…”

“…A similar strategy has been described in which adenovirus was included in complexes of DNA plus a cationic molecule (polylysine or cationic lipids), and in some cases a targeting moiety. [5][6][7][8]10,12,14 In such strategies, the adenovirus was either covalently linked to the cationic molecule or included in the complex via electrostatic interactions. Including adenovirus in the complex by either method markedly improved delivery and expression of plasmid DNA.…”