Abstract: In one aspect, the present invention provides an intron-modified capsid expression cassette useful for generating adeno-associated virus (AAV) vector particles. In another aspect, the present invention provides a method of reducing the immune response in a mammalian subject undergoing treatment with an AAV vector.

Abstract: In one aspect, the present invention provides an intron-modified capsid expression cassette useful for generating adeno-associated virus (AAV) vector particles. In another aspect, the present invention provides a method of reducing the immune response in a mammalian subject undergoing treatment with an AAV vector.

Abstract: In one aspect, the present invention provides an intron-modified cap expression cassette useful for generating adeno-associated virus (AAV) vector particles. In another aspect, the present invention provides a method of reducing the immune response in a mammalian subject undergoing treatment with an AAV vector.

Abstract: In one aspect, the present invention provides an intron-modified capsid expression cassette useful for generating adeno-associated virus (AAV) vector particles. In another aspect, the present invention provides a method of reducing the immune response in a mammalian subject undergoing treatment with an AAV vector.

Abstract: In one aspect, the present invention provides an intron-modified cap expression cassette useful for generating adeno-associated virus (AAV) vector particles. In another aspect, the present invention provides a method of reducing the immune response in a mammalian subject undergoing treatment with an AAV vector.

Abstract: The present invention provides packaging cell lines for the efficient production of an Adeno-associated virus (AAV) vector which does not require “helper” virus function for the replication and encapsidation of the AAV vector particles. Packaging cells, methods for their production and methods for producing recombinant AAV vector particles useful for human gene therapy are provided.

Abstract: The present invention provides packaging cell lines for the efficient production of an Adeno-associated virus (AAV) vector which does not require “helper” virus function for the replication and encapsidation of the AAV vector particles. Packaging cells, methods for their production and methods for producing recombinant AAV vector particles useful for human gene therapy are provided.

Abstract: The present invention demonstrates that JSRV envelope protein (Env) can be used to transduce human and other mammalian cells. Hybrid retrovirus packaging cells have been constructed that express the JSRV Env and retrovirus Gag-Pol proteins, and can produce JSRV-pseudotype vectors at high titers. Using high-titer virus the host range for JSRV has been established, and included sheep, human, monkey, bovine and dog cells, but not murine, rat or hamster cells. Retroviral packaging cell lines comprising the JSRV envelope protein, and receptor binding fragments thereof, are provided which transiently and stably produce high titers of recombinant retrovirus particles which can be used to transfer a heterologous gene to a eukaryotic cell.

Abstract: rDNA promoter constructs useful in plasmids and vectors directing transcription of RNAs in a Pol I-specific manner constructed of four elements in a serial array: a first nucleotide sequence, capable of hybridizing under stringent conditions to an rDNA promoter element; a second nucleotide sequence, capable of hybridizing under stringent conditions to an internal ribosome entry signal (IRES); a third nucleotide sequence containing a coding region interest; a fourth nucleotide sequence containing a polyadenylation (polyA) signal sequence; a fifth rDNA enhancer element may be positioned upstream of the serial array. Also recombinant permissive cells and genetically engineered stable cell lines that contain the subject constructs.

Abstract: Retroviral vectors for producing coordinately expressed polycistronic mRNA in transfected host cells. A representative retroviral construct capable of forming a proviral genome in a host cell contains a first nucleotide coding sequence, a second nucleotide coding sequence, and a third nucleotide sequence capable of hybridizing under stringent conditions to a 5′ nontranslated region (NTR) of a picornavirus RNA or its complementary RNA strand. The first, second, and third nucleotide sequences are operably linked such that transcription of the proviral genome gives rise to a messenger RNA molecule containing transcripts of the first, second, and third nucleotide sequences. The transcript of the third nucleotide sequence in the messenger RNA molecule contains a nucleic acid capable of forming a regulatory stem-loop nucleic acid structure followed by at least one operable AUG start codon.

Abstract: Retroviral packaging cells produce replication-defective retroviral particles capable of binding to Mus dunni endogenous virus retroviral receptors on target cells and are useful in gene transfer and gene therapy. The packaging cell employs a vector encoding a M. dunni retroviral Env protein and produces the retroviral particles at high titer.

Abstract: This invention includes methods for increasing the efficiency of transduction of cells, including non-dividing cells, by recombinant AAV vectors. The methods utilize agents that alter certain aspects of DNA metabolism, more specifically, that affect DNA synthesis and/or affect repair, that impact on maintenance of chromosomal integrity, and/or that cause damage to the cellular DNA. Agents and vectors can now also be preselected and screened for transducing ability and/or transducing agents for their effect on DNA metabolism. These agents include tritiated nucleotides such as thymidine, gamma irradiation, UV irradiation, cis-platinum, etoposide, hydroxyurea and aphidicolin.

Abstract: Retroviral packaging cells produce replication-defective retroviral vector particles capable of binding to Glvr-1 or Ram-1 retroviral receptors on target cells and are useful in gene therapy. The packaging cell employs a vector encoding a 10A1 retroviral env protein and produces the retroviral particles at high titer.

Abstract: The invention includes methods for increasing the efficiency of transduction of cells, including non-dividing cells, by recombinant AAV vectors. The methods utilize agents that alter certain aspects of DNA metabolism, more specifically, that affect DNA synthesis and/or affect repair, that impact on maintenance of chromosomal integrity, and/or that cause damage to the cellular DNA. Agents and vectors can now also be preselected and screened for transducing ability and/or transducing agents for their effect on DNA metabolism. These agents include tritiated nucleotides such as thymidine, gamma irradiation, UV irradiation, cis-platinum, etoposide, hydroxyurea and aphidicolin.

Abstract: Retrovirus packaging cell lines PG13 (ATCC No. CRL 10686) and PG13/LNc8 (ATCC No. CRL 10685), and retroviruses packaged by said cells.

Abstract: A process of mammalian gene therapy. Explanted fibroblasts are genetically modified by introducing a retroviral construct containing a nucleotide sequence encoding for a therapeutic substance. The genetically modified fibroblasts are implanted into a mammalian subject.

Abstract: DNA constructs consisting essentially of the promoter, gag, pol, and env sequences of a helper virus useful for making retrovirus packaging cell lines that do not yield helper virus and do not transfer the packaging function. Such DNA molecules are constructed by deleting from the genome of a replication-competent retrovirus all cis-acting elements except for the tRNA binding site. Specifically, deletion is made of the packaging signal, the site for initiation of second strand DNA synthesis, the site required for translation of reverse transcriptase during first strand DNA synthesis, and the provirus integration signal. DNA construct pPAM3 (ATCC No. 40234) is a representative embodiment. A cell line containing such an altered viral genome does not transmit this virus or transfer the packaging signal, but will transmit high titers of other viral RNAs containing the proper cis-acting elements, including retroviral vectors designed to carry foreign genes. Cell line PA317 (ATCC No.