Abstract: The present invention provides methods for administering an adenoviral gene transfer vector comprising an exogenous gene to an animal. One method involves utilizing systemic neutralizing antibodies to neutralize the adenoviral gene transfer vector outside a targeted muscle. Another method involves the repeat administration of an adenoviral gene transfer vector to a skeletal muscle.

Abstract: The present invention provides multiply deficient adenoviral vectors and complementing cell lines. Also provided are recombinants of the multiply deficient adenoviral vectors and a therapeutic method, particularly relating to gene therapy, vaccination, and the like, involving the use of such recombinants.

Abstract: The present invention provides a dual selection cassette (DSC) comprising first and second DNA segments having homology to a eukaryotic viral vector, positive and negative selection genes, each operably linked to their own promoter, and one or more unique restriction enzyme sites (URES) or sitey-directed homologous recombination sites. The present invention also provides a plasmid, pN/P, comprising an independent positive selection marker gene, an origin of replication, and a dual selection cassette. The dual selection cassette and pN/P plasmid can be used to produce eukaryotic gene transfer vectors without requiring temporally-linked double recombination events or the use of specialized bacterial strains that allow the replication of plasmids comprising defective origins of replication. This method usefully increases the ratio of desired to undesired plasmid and vector constructs. Additionally, this invention provides a method for the creation of eukaryotic viral vector libraries.

Abstract: The present invention provides a chimeric adenovirus fiber protein, which differs from the wild-type coat protein by the introduction of a nonnative amino acid sequence in a conformationally-restrained manner. Such a chimeric adenovirus fiber protein according to the invention is able to direct entry into cells of a vector comprising the chimeric fiber protein that is more efficient than entry into cells of a vector that is identical except for comprising a wild-type adenovirus fiber protein rather than the chimeric adenovirus fiber protein. The nonnative amino acid sequences encodes a peptide motif that comprises an epitope for an antibody, or a ligand for a cell surface receptor, that can be employed in cell targeting. The present invention also pertains to vectors comprising such a chimeric adenovirus fiber protein, and to methods of using such vectors.

Abstract: The present invention provides a dual selection cassette (DSC) comprising first and second DNA segments having homology to a eukaryotic viral vector, positive and negative selection genes, each operably linked to their own promoter, and one or more unique restriction enzyme sites (URES) or site-directed homologous recombination sites. The present invention also provides a plasmid, pN/P, comprising an independent positive selection marker gene, an origin of replication, and a dual selection cassette. The dual selection cassette and pN/P plasmid can be used to produce eukaryotic gene transfer vectors without requiring temporally-linked double recombination events or the use of specialized bacterial strains that allow the replication of plasmids comprising defective origins of replication. This method usefully increases the ratio of desired to undesired plasmid and vector constructs. Additionally, this invention provides a method for the creation of eukaryotic viral vector libraries.

Abstract: The present invention provides a chimeric protein IX (pIX). The chimeric pIX protein has an adenoviral pIX domain and also a non-native amino acid. Where the non-native amino acid is a ligand that binds to a substrate present on the surface cells, the chimeric pIX can be used to target vectors containing such proteins to desired cell types. Thus, the invention provides vector systems including such chimeric pIX proteins as well as methods of infecting cells using such vector systems.

Abstract: The present invention provides a library of viral vectors, wherein each member comprises a first heterologous DNA encoding a first gene product and a second heterologous DNA encoding a second gene product. The first heterologous DNA is common to each member of the library, while the second heterologous DNA varies between members of the library. The present invention additionally provides a method of constructing a library of viral vectors. The method comprises carrying out homologous recombination between a first DNA molecule and a second DNA molecule to form a pool of intermediate viral vector genomes. One or more linear third DNA molecules are ligated into the pool of intermediate viral genomes to produce a library of viral vector genomes. Alternatively, homologous recombination between linear DNA molecules and recipient DNA molecules produces a library of viral vector genomes. The library of viral vector genomes is converted into a library of viral vectors.

Abstract: The present invention provides a method of modulating the persistence of expression of a transgene in an at least E4&Dgr; adenoviral vector in a cell. In one embodiment, the method comprises contacting the cell with an at least E4&Dgr; adenoviral vector comprising (i) a transgene and (ii) a gene encoding a trans-acting factor, which is not from the E4 region of an adenovirus and which modulates the persistence of expression of the transgene. In another embodiment, the method comprises contacting the cell simultaneously or sequentially with (i) an at least E4&Dgr; adenoviral vector comprising a transgene and (ii) a viral vector comprising a gene encoding a trans-acting factor, which is not from the E4 region of an adenovirus and which modulates the persistence of expression of the transgene. In addition, the present invention provides a recombinant at least E4&Dgr; adenoviral vector for use in the method and a composition comprising the vector and a carrier therefor.

Abstract: The present invention provides a method and composition for preserving an adenoviral vector. Both the method and composition involve the use of an adenoviral gene transfer vector, a pharmaceutically acceptable liquid carrier, and a stabilizing agent, which enables the adenoviral vector to be maintained at a temperature above 0° C. for at least 7 days.

Abstract: The present invention provides a method of modulating the persistence of expression of a trans gene in an at least E4&Dgr; adenoidal vector in a cell. In one embodiment, the method comprises contacting the cell with an at least E4&Dgr; adenoidal vector comprising (i) a transgene and (ii) a gene encoding a trans-acting factor, which is not from the E4 region of an adenovirus and which modulates the persistence of expression of the transgene. In another embodiment, the method comprises contacting the cell simultaneously or sequentially with (i) an at least E4&Dgr; adenoidal vector comprising a transgene and (ii) a viral vector comprising a gene encoding a trans-acting factor, which is not from the E4 region of an adenovirus and which modulates the persistence of expression of the transgene. In addition, the present invention provides a recombinant at least E4&Dgr; adenoviral vector for use in the method and a composition comprising the vector and a carrier therefor.

Abstract: The present invention provides a method of producing an adenoviral vector stock by providing a culture of cells permissive for growth of adenoviral vectors, wherein the cells are in a medium, culturing the culture under conditions to permit growth of the cells, perfusing fresh medium through the culture for a period of about 1-6 hours, in an amount of at least about two times the volume of medium in the culture, while the density of the cells in the medium is about 40-70% of the density of cells obtained in the medium when the growth of the culture is in the stationary phase, contacting the culture with adenoviral vectors under conditions permissive for the infection of the cells after the perfusion of fresh medium is substantially completed, culturing the infected cells to replicate the adenoviral vectors, and harvesting the infected cells to obtain an adenoviral vector stock.

Abstract: The present invention provides a chimeric adenoviral coat protein (particularly a chimeric adenovirus hexon and/or fiber protein). The chimeric adenovirus coat protein has a decreased ability or inability to be recognized by a neutralizing antibody directed against the corresponding wild-type adenovirus coat protein. The invention also provides an adenovirus comprising a chimeric adenovirus coat protein, and methods of constructing and using such an adenovirus, for instance, in gene therapy.

Abstract: The present invention provides a chimeric adenoviral coat protein (particularly a chimeric adenovirus hexon and/or fiber protein). The chimeric adenovirus coat protein has a decreased ability or inability to be recognized by a neutralizing antibody directed against the corresponding wild-type adenovirus coat protein. The invention also provides an adenovirus comprising a chimeric adenovirus coat protein, and methods of constructing and using such an adenovirus, for instance, in gene therapy.

Abstract: The present invention provides a recombinant adenovirus having a genome with a deficiency in the E1 region and a mutation in the MLP. The invention also provides stocks of such viruses and compositions including the inventive viruses. The invention also provides cell lines for propagating the recombinant adenoviruses.

Abstract: The present invention provides methods for site-specific recombination in a cell, as well as vectors which can be employed in such methods. The methods and vectors of the present invention can be used to obtain persistent gene expression in a cell and to modulate gene expression.One preferred method according to the invention comprises contacting a cell with a vector comprising an origin of replication functional in mammalian cells located between first and second recombining sites located in parallel. Another preferred method comprises, in part, contacting a cell with a vector comprising first and second recombining sites in antiparallel orientations such that the vector is internalized by the cell. In both methods, the cell is further provided with a site-specific recombinase that effects recombination between the first and second recombining sites of the vector.

Abstract: The present invention provides a chimeric adenovirus fiber protein, which differs from the wild-type coat protein by the introduction of a nonnative amino acid sequence in a conformationally-restrained manner. Such a chimeric adenovirus fiber protein according to the invention is able to direct entry into cells of a vector comprising the chimeric fiber protein that is more efficient than entry into cells of a vector that is identical except for comprising a wild-type adenovirus fiber protein rather than the chimeric adenovirus fiber protein. The nonnative amino acid sequences encodes a peptide motif that comprises an epitope for an antibody, or a ligand for a cell surface receptor, that can be employed in cell targeting. The present invention also pertains to vectors comprising such a chimeric adenovirus fiber protein, and to methods of using such vectors.

Abstract: The present invention provides multiply deficient adenoviral vectors and complementing cell lines. Also provided are recombinants of the multiply deficient adenoviral vectors and a therapeutic method, particularly relating to gene therapy, vaccination, and the like, involving the use of such recombinants.

Abstract: The present method involves evaluating the purity of a viral gene transfer vector stock by providing an aliquot of the viral gene transfer vector stock and producing charged fragments from the aliquot. The fragments are detected by a detector that produces a sample signal corresponding to the mass-to-charge ratio of the detected fragments. This signal then is compared to a standard signal, whereby the purity of the viral gene transfer vector stock is evaluated.

Abstract: The present invention provides a chimeric adenovirus coat protein, which differs from the wild-type coat protein by the introduction of a nonnative amino acid sequence. Such a chimeric adenovirus coat protein according to the invention is able to direct entry into cells of a vector comprising the coat protein that is more efficient than entry into cells of a vector that is identical except for comprising a wild-type adenovirus coat protein rather than the chimeric adenovirus coat protein. The chimeric coat protein preferably is a fiber, hexon, or penton protein. The present invention also provides an adenoviral vector that comprises the chimeric adenovirus coat protein, as well as methods of constructing and using such a vector.

Abstract: The present invention provides a short-shafted adenoviral fiber, a recombinant adenovirus comprising a short-shafted fiber, a vector comprising a short-shafted adenoviral fiber gene, and a method of targeting attachment of a short-shafted recombinant adenovirus to a cell so as to effect entry of the short-shafted recombinant adenovirus into the cell.