Abstract: Methods of assembling modified adenoviruses, libraries of adenoviral gene modules and compositions thereof are provided herein.

Abstract: Described herein are viral amplicon-based protein expression systems and methods useful for producing heterologous proteins, such as enzymes, by agroinfiltration. The methods involve producing an Agrobacterium with a Ti plasmid encoding a heterologous protein, infecting plant cells with the Agrobacterium, allowing expression of the heterologous protein, and recovering the heterologous protein from the plant cells. In one embodiment, the protein produced is an endoglucanase.

Abstract: This disclosure provides for compositions and methods for the use of designed nucleic acid-targeting nucleic acids, Argonautes, and complexes thereof.

Abstract: Methods for producing reassortant viruses are provided wherein the transcription and/or translation of the hemagglutinin and/or neuraminidase genes are suppressed.

Abstract: Disclosed herein are methods and compositions for targeted integration of one or more copies of a sequence of interest using zinc finger nucleases (ZFNs) comprising a zinc finger protein and a cleavage domain or cleavage half-domain and integrase defective lentiviral donor constructs.

Abstract: The present invention is based on the in vivo demonstration that RSV can be inhibited through intranasal administration of iRNA agents as well as by parenteral administration of such agents. Further, it is shown that effective viral reduction can be achieved with more than one virus being treated concurrently. Based on these findings, the present invention provides general and specific compositions and methods that are useful in reducing RSV mRNA levels, RSV protein levels and viral titers in a subject, e.g., a mammal, such as a human. These findings can be applied to other respiratory viruses.

Abstract: There is provided an HSV complex which comprises an avirulent HSV and a targeting agent which allows the HSV particle to infect and lyse a specific targeted cell. The inventors have found a way in which avirulent HSV can be targeted to disease cells, e.g. cancer cells, by incorporating an antibody binding domain into one or more viral glycoproteins.

Abstract: The present invention relates to the NQBS class of molecules. It is based, at least in part, on the discovery that a representative group of compounds have been observed to inhibit nuclear translocation of NF-?B subunits. Without being bound by any particular theory, this inhibition of nuclear translocation may be mediated by either (i) binding of the NQBS or related compound to the C-terminus of the RHD, which specifically mediates the nuclear internalization; or (ii) NQBS-mediated stabilization of the dimer/I?B complex, disallowing dissociation of the active NF-?B monomers, and thus, inhibiting the generation of the subunits necessary to enter the nucleus. The NQBS class of molecules, and related molecules, may be used in therapeutic applications where inhibition of NF-?B translocation is beneficial, including but not limited to the treatment of cancer, autoimmune disorders, and inflammatory states.

Abstract: It is intended to provide a polynucleotide comprising a viral base sequence, the viral base sequence containing: a first base sequence encoding a viral replication protein, and a second base sequence encoding a viral movement protein, the second base sequence being located downstream of the first base sequence and having a linking site for linking with an exogenous base sequence encoding a polypeptide to be expressed, the linking site being located downstream of the second base sequence, the second base sequence being obtained by modifying with a base sequence in a native sequence derived from a virus by insertion, substitution, or addition. By using this, a vector containing a viral base sequence is constructed, and a protein is efficiently produced without worsening growth of a host cell containing the vector.

Abstract: Lambda phages that can be used to introduce recombineering functions into host cells are disclosed. Also disclosed are plasmids that can be used to confer recombineering functions to a variety of strains of E. coli and to other bacteria, including Salmonella, Pseudomonas, Cyanobacteria, Spirochaetes. These plasmids and phages can be isolated in vitro and can be used to transform bacterial cells, such as gram negative bacteria.

Abstract: TRPC ion channels are non-selective channels widely expressed in human tissues. These channels are involved in numerous physiological functions and are putative targets for the development of novel medicines. There is a need to gain a better understanding of TRPC ion channels in cells and beyond. The present invention provides a pharmacological tool compound that allows to study TRPC ion channels due to its discriminating inhibition of TRPC subfamilies. In the present invention, norgestimate is shown to selectively inhibit TRPC3, TRPC6 and TRPC7.

Abstract: The invention is related to a dengue virus or chimeric dengue virus that contains a mutation in the 3? untranslated region (3?-UTR) comprising a ?30 mutation that removes the TL-2 homologous structure in each of the dengue virus serotypes 1, 2, 3, and 4, and nucleotides additional to the ?30 mutation deleted from the 3?-UTR that removes sequence in the 5? direction as far as the 5? boundary of the TL-3 homologous structure in each of the dengue virus serotypes 1, 2, 3, and 4, or a replacement of the 3?-UTR of a dengue virus of a first serotype with the 3?-UTR of a dengue virus of a second serotype, optionally containing the ?30 mutation and nucleotides additional to the ?30 mutation deleted from the 3?-UTR; and immunogenic compositions, methods of inducing an immune response, and methods of producing a dengue virus or chimeric dengue virus.

Abstract: This invention provides methods for obtaining specific and stable integration of nucleic acids into eukaryotic cells. The invention makes use of site-specific recombination systems that use prokaryotic recombinase polypeptides, such as the ?C31 integrase, that can mediate recombination between the recombination sites, but not between hybrid recombination sites that are formed upon the recombination. Thus, the recombination is irreversible in the absence of additional factors. Eukaryotic cells that contain the recombinase polypeptides, or genes that encode the recombinases, are also provided.

Abstract: The instant invention provides methods and compositions for generating recombinant adenoviral vectors. The invention also provides kits comprising for the generation of recombinant adenoviral vectors.

Abstract: A polypeptide comprising a preS1 region of hepatitis B virus (HBV), or a fragment thereof, and/or the preS2 region of HBV or a fragment thereof, and methods of use to inhibit virus infection are disclosed. A lentivirus comprising hepatitis B virus (HBV) envelope proteins, or a fragment thereof, and/or the L envelope protein of HBV and/or the M envelope protein of HBV or a fragment thereof, and/or the S envelope protein of HBV or a fragment thereof, and methods of use of this lentivirus HBV pseudovirus as a gene therapy to target hepatocytes for the administration of therapeutic agents are also disclosed.

Abstract: There is provided an HSV complex which comprises an avirulent HSV and a targeting agent which allows the HSV particle to infect and lyse a specific targeted cell. The inventors have found a way in which avirulent HSV can be targeted to disease cells, e.g. cancer cells, by incorporating an antibody binding domain into one or more viral glycoproteins.

Abstract: The present invention provides methods and compositions related to modulating the resistance of a cell against a target nucleic acid or a transcription product thereof. In some preferred embodiments, the present invention provides compositions and methods for the use of one or more cas genes or proteins for modulating the resistance of a cell against a target nucleic acid or a transcription product thereof. In some embodiments, the present invention provides methods and compositions that find use in the development and use of strain combinations and starter culture rotations. In additional embodiments, the present invention provides methods for labelling and/or identifying bacteria. In some preferred embodiments, the present invention provides methods for the use of CRISPR loci to determine the potential virulence of a phage against a cell and the use of CRISPR-cas to modulate the genetic sequence of a phage for increased virulence level.

Abstract: In one aspect, the invention provides methods and compositions for the expression of small RNA molecules within a cell using a retroviral vector (FIG. 1A). The methods can be used to express double stranded RNA complexes. Small interfering RNA (siRNA) can be expressed using the methods of the invention within a cell, that interfere with a viral life cycle by down regulating either the viral genome, a viral genome transcript, or a host cell that. In another aspect the invention provides methods for treating patients having suffering from infection, particularly infection with HIV. In a further aspect, the invention provides methods for producing siRNA encoding lentivirus where the siRNA activity may interfere with the lentiviral life cycle.

Abstract: For a method of inducing differentiation of cardiomyocytes from stem cells, a method is provided to induce efficiently and selectively differentiation of cardiomyocytes by such a method in which the stem cells are cultured to induce differentiation into cardiomyocytes in the presence of a substance that inhibits BMP signaling.

Abstract: A novel system, including vectors, phage particles, host cells and methods of use, for expressing one or more polypeptides of interest in prokaryotes is described.

Abstract: The invention provides novel methods of gene targeting using replication in order to increase the efficiency of targeted genetic modification in an eukaryotic organism. Included are vectors, expression cassettes, and modified cells, plants and seeds.

Abstract: Live, attenuated, phenotypically stable HSV-2 viruses and methods of making and using the virus are provided. Live, attenuated HSV-2 viruses are constructed using recombinant techniques and can be used in a pharmaceutical composition for prophylactic treatment of HSV-2 infections and for treatment of recurrent HSV-2 related diseases and conditions.

Abstract: The invention belongs to the field of animal health, in particular equine diseases caused by equine herpesvirus (EHV). The invention relates to artificial chromosomes comprising the genome of equine herpesviruses, methods of producing attenuated or virulent EHV with or without the insertion of foreign genes, EHV obtainable with said methods and pharmaceutical compositions comprising said viruses.

Abstract: Regarding a card storage cassette for use in a card recording apparatus in which a plurality of cards are stacked vertically with their face maintained substantially horizontal, the present invention provides in one embodiment a card recording apparatus and its card storage cassette in which cards can be replenished even when the undermost card is being ejected from the card storage cassette. While the card storage cassette (30) to be detachably mounted on the cassette mount platform formed on the right side plate of the outer chassis of the card recording apparatus is framed by joining a cassette body and a cassette cover, the cassette cover can be removed from the cassette body along a vertical guide member.

Abstract: A lytic virus specific for a target strain of a microorganism and substantially free of undesirable genes may be utilized in processes including control of populations of microorganisms. The virus may include a host-range mutant, or “h-mutant.” A method for generating virus includes growing virus-resistant variants of a target strain of a microorganism in the presence of viruses that are specific for the target strain. Only h-mutant viruses will proliferate. Wild-type virus-resistant and virus-resistant variants of a microorganism are also disclosed, as are methods generating such variants. Methods for controlling target strain microorganisms include introducing virus into a treatment site where control of a population of a target strain microorganism is desired or introducing virus-resistant variants of a microorganism into treatment sites where the presence of the microorganism is desired.

Abstract: This disclosure provides a system for producing pancreatic islet cells from embryonic stem cells. Differentiation is initiated towards endoderm cells, and focused using reagents that promote emergence of islet precursors and mature insulin-secreting cells. High quality populations of islet cells can be produced in commercial quantities for use in research, drug screening, or regenerative medicine.

Abstract: In one aspect, the invention provides methods and compositions for the expression of small RNA molecules within a cell using a retroviral vector. The methods can be used to express double stranded RNA complexes. Small interfering RNA (siRNA) can be expressed using the methods of the invention within a cell, that interfere with a viral life cycle by down regulating either the viral genome, a viral genome transcript, or a host cell that. In another aspect the invention provides methods for treating patients having suffering from infection, particularly infection with HIV. In a further aspect, the invention provides methods for producing siRNA encoding lentivirus where the siRNA activity may interfere with the lentiviral life cycle.

Abstract: The present invention provides an advance in phage display technology by permitting the uncoupling of the propagation of phages containing inserted sequences encoding heterologous polypeptides from the expression of said polypeptides. The invention provides phage constructs and methods for their use to permit phage coat protein expression, and thus phage propagation, in the absence of display of heterologous polypeptides, which may be expressed as a fusion with said coat protein in a regulated manner.

Abstract: Methods are provided for the rapid identification of essential or conditionally essential DNA segments in any species of haploid cell (one copy chromosome per cell) that is capable of being transformed by artificial means and is capable of undergoing DNA recombination. This system offers an enhanced means of identifying essential function genes in diploid pathogens, such as gram-negative and gram-positive bacteria.

Abstract: The present invention is based on the development of a dual promoter system (preferably a RNA pol I-pol II system) for the efficient intracellular synthesis of viral RNA. The resultant minimal plasmid-based system may be used to synthesize any RNA virus, preferably viruses with a negative single stranded RNA genome. The viral product of the system is produced when the plasmids of the system are introduced into a suitable host cell. One application of the system is production of attenuated, reassortant influenza viruses for use as antigens in vaccines. The reassortant viruses generated by cotransfection of plasmids may comprise genes encoding the surface glycoproteins hemagglutinin and neuramimidase from an influenza virus currently infecting the population and the internal genes from an attenuated influenza virus. An advantageous property of the present invention is its versatility; the system may be quickly and easily adapted to synthesize an attenuated version of any RNA virus.

Abstract: Compounds are provided that inhibit the interaction of an IGF with any one of its binding proteins and not to a human IGF receptor. These IGF agonist compounds, which include peptides, are useful to increase serum and tissue levels of active IGFs in a mammal.

Abstract: The invention provides novel replication deficient adenovirus vectors and methods for making and using these viruses. The invention also provides vector systems and kits using a serotype specific strategy for making adenoviral vector preparations substantially free of replication competent “helper” virus. The helper virus-free preparations provide novel pharmaceutical compositions substantially free of helper virus for use in gene transfer and gene therapy.

Abstract: The present invention relates to a method for producing a recombinant adeno-associated virus (rAAV), in which at different times a helper construct and also a vector construct are introduced into a suitable cell, and preferably the helper construct does not comprise, in particular with the exception of the AAV promoters, any nucleic acid sequences to which at least a Rep protein can essentially specifically bind and preferably the vector construct comprises ITR sequences in flop orientation. The recombinant adeno-associated viruses produced according to the method of the invention are suitable in particular for producing a tumor cell into which additionally nucleic acids coding for GM-CSF and B7.2 has been introduced, which in turn can be used in the form of a medicament for the treatment of cancers.

Abstract: The invention provides a novel Adenovirus backbone plasmid, which when co-transfected with a shuttle vector, allows for production of recombinant viruses quickly and easily. The present invention also provides host cells and a cloning system for generating recombinant adenoviruses.

Abstract: This invention relates to the screening of nucleic acids. More particularly, the present invention provides a dysfunctional viral genome capable of both expressing libraries of exogenous nucleic acids and selecting the sequences having a predefined characteristic or function within the cell, such as nucleic acids encoding signal peptides, secreted proteins, membrane bound proteins, proteases and drug-resistance proteins. The invention further provides a method and a kit for selecting nucleic acids having a desired feature, wherein production of a viral particle is dependent on insertion of an exogenous nucleic acid having the desired feature into a dysfunctional viral genome or into a viral genome exposed to a substance inhibiting viral packaging function(s).

Abstract: The present invention relates to (a) methods for improving a genetic stability of an insert nucleotide sequence in a recombinant single-stranded RNA virus vector, which comprises performing a mutagenesis of the foreign insert nucleotide sequence to provide even distribution of G/C content throughout the overall foreign insert nucleotide sequence and/or to increase G/C content of the foreign insert without substantially causing amino acids substitutions (b) a recombinant single-stranded RNA virus comprising an insert nucleotide sequence with improved genetic stability and (c) a recombinant poliovirus comprising an insert nucleotide sequence with improved genetic stability.

Abstract: The present invention provides a method of increasing expression of a promoter distal gene in a virus of the order Mononegavirales, and a recombinant virus constructed by such method. Also provided is a method of attenuating a virus of the order Mononegavirales, and of constructing an attenuated virus useful for a vaccine.

Abstract: The present invention is directed to isolated transducing phages, methods of isolating transducing phages, and methods of using transducing phages including, for instance, transferring at least one nucleic acid fragment from a donor microbe to a recipient microbe, and producing a secondary metabolite from a microbe. The transducing phages typically have a broad host range, and transduce microbes in the Order Actinomycetales, in particular in the Family Streptomycetaceae, including Streptomyces coelicolor, Streptomyces lividans, Streptomyces venezuelae, Streptomyces avermitilis, and Saccharopolyspora erythraea. The transducing phages can be specialized transducing phages or generalized transducing phages.

Abstract: The present invention provides a Hepatitis C Virus (HCV) replicon that efficiently replicates in an eukaryotic cell. The HCV replicon includes a nucleic acid sequence encoding a subgenomic fragments of HCV of any genotype that confer on the RNA the ability to replicate, and a nucleic acid sequence encoding an acetyl transferase selectable marker, such as puromycin. Also provided is an HCV type 1a replicon that efficiently replicates in an eukaryotic cell and includes a nucleic acid sequence encoding subgenomic fragments of type 1a HCV that confer on the RNA the ability to replicate, and a nucleic acid sequence encoding a acetyl transferase selectable marker. Further provided are eukaryotic cell lines that include an HCV replicon or an HCV type 1a replicon which efficiently replicate in the eukaryotic cell. The present invention also provides screening methods for identifying candidate compounds that inhibit the propagation of HCV.

Abstract: A method for generating and amplifying closed circular DNA having a specific sequence in vitro in a cell-free system is disclosed. Prior to the invention of this method, closed circular DNA could only be amplified in vivo in appropriate host cells. The essence of the method is the inclusion of a thermostable DNA ligase in a PCR reaction. This procedure is referred to as ligation-during-amplification (LDA), in which the fully extended DNA strands are ligated by the DNA ligase and used as templates for subsequent amplification. Closed circular DNA having a specific sequence can be selectively amplified exponentially by the use of two sequence-specific primers in the LDA reaction. In addition, one or more site-specific mutations can be introduced into a closed circular DNA by the use of one or more mutagenic primers in the LDA reaction. Various thermostable DNA polymerases and thermostable ligases can be used for LDA amplification.

Abstract: The present invention provides an improved method of making eukaryotic gene transfer vectors comprising homologous recombining lambdid vectors with a second DNA in a bacterium to generate novel recombinant eukaryotic viral gene transfer vectors as well as a novel lambdid vector used in the inventive method and an inventive system comprising the novel lambdid vector.

Abstract: The present invention is directed to isolated transducing phages, methods of isolating transducing phages, and methods of using transducing phages including, for instance, transferring at least one nucleic acid fragment from a donor microbe to a recipient microbe, and producing a secondary metabolite from a microbe. The transducing phages typically have a broad host range, and transduce microbes in the Order Actinomycetales, in particular in the Family Streptomycetaceae, including Streptomyces coelicolor, Streptomyces lividans, Streptomyces venezuelae, Streptomyces avermitilis, and Saccharopolyspora erythraea. The transducing phages can be specialized transducing phages or generalized transducing phages.

Abstract: A method for improved plastid transformation efficiency via homologous recombination and nucleic acid sequences useful therefore is provided. Nucleic acid sequences comprising a 5 base pair recombination sequence motif or multiple direct repeats thereof that increase the frequency of integration of a selected transgene through plastid transformation by homologous recombination are provided.

Abstract: An isolated DNA having the promoter sequence of the hex gene of P. chrysogenum or a DNA fragment that is hybridizable to the complement of the promoter sequence under stringent conditions and is capable of directing expression of DNA downstream of the fragment in P. chrysogenum. Also a process for promoting expression of a coding sequence of interest in a microorganism using the isolated DNA and a process to block expression of a gene of interest in a microorganism using the isolated DNA are disclosed.

Abstract: A viral DNA construct, and virus encoded thereby, is provided having one or more tumor specific transcription factor binding sites in place of one or more wild type transcription factor binding sites operatively positioned in the promoter region which controls expression of early genes responsible for viral nucleic acid replication. Preferred constructs place the tumor specific transcription factor binding sites in operative relation to DNA polymerase, DNA terminal protein and/or DNA binding protein. Compositions and constructs contained therein are provided, particularly for use in therapy. Methods of treating patients for neoplasms are also provided.

Abstract: An innovative detection system for detecting small numbers of target analytes is disclosed. This system provides a novel method for attaching multiple copies of reporter groups to a single site on an analyte of interest. This system preferably comprises a virus capsid enclosing multiple detectable reporter groups, and a linking molecule which is capable of linking the capsid to the analyte of interest.

Abstract: The gene of human acidic fibroblast growth factor 155 (haFGF 155) has been obtained by chemical synthesis. The nucleotide sequence of haFGF 155 gene has been deduced on the basis of haFGF 155 amino acid sequence as described in the literature. The amino acid sequence of the synthesized haFGF 155 does not differ from those described in the literature. The nucleotide sequence of haFGF gene differs from those described previously. For chemical synthesis of haFGF 155 gene, codons were used which are the ones most often used by E. coli in highly expressed E. coli proteins. A plasmid with haFGF 155 (phaFGF 155) gene was obtained and was used to transform E. coli. Production of haFGF 154 protein was achieved by cultivation of the producer strain under conditions which slow down the lytic development of lambda phage. The haFGF 154 protein accumulated in culture medium in a soluble condition as a result of the producer strain cells lysis by the lambda phage.

Abstract: a method is provided for detecting a presence of HIV virus in a sample comprising: taking a culture of recombinant cells which (a) are capable of cell division, (b) express CD4 receptor and one or more additional cell surface receptors necessary to allow the HIV virus to infect, (c) enable the HIV virus to replicate and infect the noninfected cells in the cell culture, and (d) comprise a reporter sequence introduced into the recombinant cells comprising a reporter gene whose expression is regulated by a protein specific to HIV viruses which is expressed from a genome of an HIV virus upon infection of the recombinant cell by the HIV virus; contacting the cell culture with a sample to be analyzed for the presence of HIV virus in the sample; and detecting a change in a level of expression of the reporter gene in cells in the recombinant cell culture.

Abstract: The present invention is directed to methods and compositions for use of homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. One aspect of the present invention relates to methods for use of bacterial conjugative transfer and homologous recombination for directed evolution, gene reassembly, and directed mutagenesis. Another aspect of the present invention relates to compositions for use in or produced by the methods of the present invention, including libraries, archives and databases.

Abstract: Disclosed are methods for inhibiting angiogenesis using cyclin dependent kinase inhibitors (CDKi) and fusion proteins thereof, recombinant viruses comprising transgenes and nucleic acid sequences encoding the same, and liposomes carrying the same as angiogenesis-inhibiting reagents.