Abstract: Non-human mammalian hosts are provided, comprising functional human hepatocytes. Isolated human hepatocytes or fragments of human hepatic tissue are introduced into the xenogeneic host in conjunction with one or more agent that stimulates human hepatocyte growth factor receptor. The human hepatocytes are maintained in the host by administration of one or more agent that stimulates human hepatocyte growth factor receptor, either continuously (e.g., via an implanted catheter or intravenous apparatus) or in discrete, regular dosages of the agent (e.g., via intravenous injections or oral dosages). The human hepatocytes are able to survive and function in the host animal for a period of at least 5 months.

Abstract: Methods and compositions for introducing a nucleic acid into the genome of at least one cell of a multicellular organism are provided. In the subject methods, a Sleeping Beauty transposon that includes the nucleic acid is administered to the multicellular organism along with a source of a Sleeping Beauty transposase activity. Administration of the transposon and transposase results in integration of the transposon, as well as the nucleic acid present therein, into the genome of at least one cell of the multicellular organism The subject methods find use in a variety of different applications, including the in vivo transfer of genes for use in, among other applications, gene therapy applications.

Abstract: Methods and compositions are provided for modulating, e.g., reducing, coding sequence expression in mammals. In the subject methods, an effective amount of an RNAi agent, e.g., an interfering ribonucleic acid (such as an siRNA or shRNA) or a transcription template thereof, e.g., a DNA encoding an shRNA, is administered to a non-embryonic mammal, e.g., via a hydrodynamic administration protocol. Also provided are RNAi agent pharmaceutical preparations for use in the subject methods. The subject methods and compositions find use in a variety of different applications, including academic and therapeutic applications.

Abstract: Methods and compositions are provided for modulating, e.g., reducing, viral coding sequence expression in mammals and mammalian cells. In the subject methods, an effective amount of an RNAi agent, e.g., an interfering ribonucleic acid (such as an siRNA or shRNA) or a transcription template thereof, e.g., a DNA encoding an shRNA, is introduced into a target cell, e.g., by being administered to a mammal that includes the target cell, e.g., via a hydrodynamic administration protocol. Also provided are RNAi agent pharmaceutical preparations for use in the subject methods. The subject methods and compositions find use in a variety of different applications, including academic and therapeutic applications.

Abstract: A helper dependent adenoviral vector system is provided. The subject helper dependent adenoviral vector system is made up of: (1) a “gutless” adenoviral vector that include cis-acting human stuffer DNA that provides for in vivo long term, high level expression of a coding sequence present on the vector; (2) an adenoviral helper vector that is characterized by having an adenoviral genome region flanked by recombinase recognition sites, where the helper vectors further include a non-mammalian endonuclease recognition site positioned outside of the adenoviral genome region; and (3) a mammalian cell that expresses the corresponding recombinase and endonuclease, as well as the adenoviral preterminal and polymerase proteins. Also provided are methods of using the subject systems to produce virions having the subject helper dependent adenoviral vectors encapsulated in an adenoviral capsid. In addition, kits for use in practicing the subject methods are provided.

Abstract: The instant invention provides methods of expressing polynucleotides in the cells of the liver comprising administering viral particles comprising a recombinant AAV vector into a mammal, preferably a human.

Abstract: Methods and compositions for introducing a nucleic acid into the genome of at least one cell of a multicellular organism are provided. In the subject methods, a Sleeping Beauty transposon that includes the nucleic acid is administered to the multicellular organism along with a source of a Sleeping Beauty transposase activity. Administration of the transposon and transposase results in integration of the transposon, as well as the nucleic acid present therein, into the genome of at least one cell of the multicellular organism The subject methods find use in a variety of different applications, including the in vivo transfer of genes for use in, among other applications, gene therapy applications.

Abstract: In one aspect, the present invention provides nucleic acid expression cassettes that are predominantly expressed in the mammalian liver. The present invention also provides vectors comprising a nucleic acid expression cassette that is predominantly expressed in the mammalian liver. The present invention also provides methods of ameliorating the symptoms of a disease, the methods including the steps of introducing into the liver of a mammalian subject a vector comprising a nucleic acid expression cassette that encodes a polypeptide, and expressing a therapeutic amount of the polypeptide in the liver.

Abstract: Methods and compositions are provided for introducing an expression cassette into a cell. In the subject methods, a population of at least two distinct adeno-associated viral particles is provided, where each distinct type of viral particle in the population comprises a different portion of the expression cassette to be introduced into the cell. The target cell is contacted with population of adeno-associated viral vectors under conditions sufficient to produce a hetero-concatemer in the cell, where the hetero-concatemer includes a functional expression cassette having an intron that includes an ITR sequence. Also provided by the subject invention are vector preparations for practicing the subject methods as well as kits for use in producing the vectors employed in the subject methods. The subject methods find use in a variety of different gene transfer applications, including both in vivo and in vitro gene transfer applications, and are particularly suited for use in the transfer of long genes.

Abstract: In vitro methods for making a recombinant adenoviral genome, as well as kits for practicing the same and the recombinant adenovirus vectors produced thereby, are provided. In the subject methods, the subject genomes are prepared from first and second vectors. The first vector includes an adenoviral genome having an E region deletion and three different, non-adenoviral restriction endonuclease sites located in the E region. The second vector is a shuttle vector and includes an insertion nucleic acid flanked by two of the three different non-adenoviral restriction endonucleases sites present in the first vector. Cleavage products are prepared from the first and second vectors using the appropriate restriction endonucleases. The resultant cleavage products are then ligated to produce the subject recombinant adenovirus genome. The subject adenoviral genomes find use in a variety of application, including as vectors for use in a variety of applications, including gene therapy.

Abstract: The invention provides compositions of a non-adenoviral vector containing a polynucleotide sequence encoding adenoviral pTP operationally linked domain. The invention also provides compositions of an adenoviral pTP binding domain. The invention also provides methods for increasing the expression of a polynucleotide by expressing the polynucleotide in a non-adenoviral vector containing an adenoviral pTP binding domain in the presence of adenoviral pTP. The invention additionally provides methods to increase expression of a heterologous polynucleotide in an individual by obtaining cells from the individual, genetically altering the cells to express a non-adenoviral vector containing an adenoviral pTP binding domain and a gene encoding pTP and readministering the genetically altered cells to the individual.

Abstract: Adenoviral vectors are used for high efficiency transduction of ribozymes specific for hepatitis C virus RNA. Hepatocytes are transduced with a recombinant adenovirus vector that expresses a ribozyme capable of specifically cleaving HCV RNA. The compositions and methods thus provide new means for treating HCV, and further provide transgenic non-human animals having human hepatocytes which are useful in models of HCV disease for developing therapeutic and preventative agents.

Abstract: A method of inhibiting hepatitis C virus RNA replication or expression is provided. The method consists of introducing two or more ribozymes specific for hepatitis C virus RNA into a cell infected with hepatitis C virus. The ribozymes specific for hepatitis C virus RNA can specifically cleave hepatitis C RNA in a HCV 5' non-coding sequence, the capsid sequence, the NS-5 sequence or any other conserved region of the hepatitis C RNA. The ribozymes can also be selected so as to be specific for opposite strands of the virus genome. A method of inhibiting hepatitis C virus RNA replication or expression is also provided which consists of introducing into a cell infected with hepatitis C virus at least one ribozyme specific for hepatitis C virus which is selected from the group consisting of GGGAGGTCTCGTAGA [SEQ ID NO: 1], GCACCATGAGCACGA [SEQ ID NO: 2], CCCACAGGACGTCAA [SEQ ID NO: 3], CAACCGTCGCCCACA [SEQ ID NO: 4], TAAACCTCAAAGAAA [SEQ ID NO: 5] GTAAGGTCATCGATA [SEQ ID NO: 6].

Abstract: The invention provides a method for enhancing the expression of a gene of interest by a cell, the cell (a) comprises a recombinant nucleic acid sequence encoding and (b) is capable of expressing the gene of interest, the method comprising contacting the cell with an amount of a soluble CTLA4 molecule effective to enhance the expression of the gene of interest by the cell.

Abstract: A combination of retroviral and adenoviral vectors are used for high efficiency gene transfer into hepatocytes, resulting in long term gene expression. Hepatocytes are transduced in vivo with a recombinant adenovirus vector that expresses a molecule capable of inducing hepatocyte regeneration, such as urokinase plasminogen activator (uPA) or tissue plasminogen activator (tPA), resulting in a high rate of liver regeneration. During the regenerative phase, ex vivo or in vivo retroviral-mediated gene transfer into hepatocytes results in greater transduction efficiencies. The compositions and methods thus provide new means for gene therapy, and transgenic non-human animals useful in developing new therapeutic and preventative agents.

Abstract: A gunsight comprises a unitary frame (2) for securing to a gun. The frame (2) has a ring (20) with a lens (22) at its front end. A light source (36) is secured to the frame rear end to project a light spot onto the lens (22). The lens (22) and the light source (36) may be independently adjustable in the vertical and horizontal planes, respectively. Alternatively, where the light source (36) is fixed the lens (22) may be adjusted in the vertical and horizontal planes. The light source (36) and the lens (22) are positioned relative to each other and to the gun barrel axis such that the light spot projected onto the lens (22) is reflected to a shooter's eye who then lines up the light spot onto a target to take aim.

Abstract: A rock bit is provided that comprises a generally cylindrical body with a leading end for boring in a rock formation and a trailing end for connection to a drill rod. The body has a generally cylindrical outside surface and comprises a plurality of parallel longitudinal grooves circumferentially spaced around the outside surface and extending from the leading end to the trailing end. The grooves have a depth that increases from the leading end to the trailing end such that the cross-sectional area of the grooves also increases towards the trailing end of the body of the bit. The body also comprises at least one channel machined in the outside surface of the body that extends between adjacent pairs of longitudinal grooves at an oblique angle to the grooves.