Abstract: The present invention provides a method for at least in part decreasing the production of an aberrant protein in a cell, the cell comprising pre-mRNA comprising exons coding for the protein, by inducing so-called exon skipping in the cell. Exon-skipping results in mature mRNA that does not contain the skipped exon, which leads to an altered product of the exon codes for amino acids. Exon skipping is performed by providing a cell with an agent capable of specifically inhibiting an exon inclusion signal, for instance, an exon recognition sequence, of the exon. The exon inclusion signal can be interfered with by a nucleic acid comprising complementarity to a part of the exon. The nucleic acid, which is also herewith provided, can be used for the preparation of a medicament, for instance, for the treatment of an inherited disease.

Abstract: The present invention provides methods and compositions for reprogramming somatic cells to a more primitive state, such as induced pluripotent stem cells, using homologous recombination. The induced pluripotent stem cells generated by the methods of the present invention are useful in a variety of therapeutic applications in the treatment and prevention of diseases and disorders.

Abstract: Aspects featured in the invention relate to compositions and methods for inhibiting alpha-synuclein (SNCA) gene expression, such as for the treatment of neurodegenerative disorders. An anti-SNCA agent featured herein that targets the SNCA gene can have been modified to alter distribution in favor of neural cells.

Abstract: Methods and kits for identifying novel anti-tumor agents are provided.

Abstract: We disclose a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a desired replacement sequence, comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing a DNA molecule comprising a first DNA fragment comprising a desired replacement sequence flanked at its 5? and 3? sides by DNA sequences substantially homologous to sequences of the chromosomal DNA flanking the target sequence and a second DNA fragment comprising an expression cassette comprising a gene encoding a selection marker operably linked to regulatory sequences functional in the eukaryotic cell, transforming the modified eukaryotic cells with the DNA molecule, selecting transformed progeny cells having the DNA molecule inserted into the chromosome, deselecting transformed progeny cells having the DNA molecule inserted into the chromosome via NHR by expression of the selection mar

Abstract: The invention provides cloned transgenic ungulates (e.g., bovines) in which prion protein activity is reduced by one or more genetically engineered mutations. Desirably, these transgenic bovines are also genetically modified to express xenogenous (e.g., human) antibodies. Because of their resistance to prion-related diseases such as bovine spongiform encephalopy (also known as mad cow disease), these bovines are a safer source of human antibodies for pharmaceutical uses and a safer source of agricultural products.

Abstract: The present invention relates to methods and compositions for controlling the expression of a target gene, whereby an intron cassette such as INT9, an intronic mec-2-derived element, is incorporated into the target gene and expression of the product of the target gene is conditional upon functional expression of the RNA processing protein, mec-8.

Abstract: The present invention relates to a method of expressing an objective protein at a high level and stably as well as for a long period even in the absence of a selection drug with a recombinant mammal cell. More particularly, the present invention relates to a method of producing an objective protein by providing a recombinant mammal cell having multiple copies of the exogenous objective protein gene expression unit integrated into a hypoxanthine-phosphoribosyl transferase enzyme (hprt) gene locus and culturing said cell.

Abstract: A new type of gene trap cassette, which can induce conditional mutations, relies on directional site-specific recombination systems, which can repair and re-induce gene trap mutations when activated in succession. After the gene trap cassettes are inserted into the genome of the target organism, mutations can be activated at a particular time and place in somatic cells. The gene trap cassettes also create multipurpose alleles amendable to a wide range of post-insertional modifications. Such gene trap cassettes can be used to mutationally inactivate all cellular genes temporally and/or spatially. Cells which contain the inventive gene trap cassette can be used for identification and/or isolation of genes and for the creation of transgenic organisms to study gene function at various developmental stages, including the adult, as well as for the creation of animal models of human disease useful for in vivo drug target validation.

Abstract: A Clostridium thermocellum thermostable cellulase enzyme with both endocellulase activity and exocellulase activity that is able to degrade cellulose in the absence of scaffolding and other cellulosomic proteins is provided. The use of the enzyme to degrade cellulosic materials to soluble sugars is also provided.

Abstract: The present invention relates to a biological entity, notably a rat, carrying a regulator construct comprising a specific repressor gene and a responder construct comprising at least one segment corresponding to a short hairpin RNA (shRNA) or corresponding to complementary short interfering RNA (siRNA) strands or corresponding to miRNA, said at least one segment being under control of a promoter which contains an operator sequence corresponding to the repressor. The invention further relates to a method for preparing said biological entity and its use.

Abstract: The present invention provides a method of achieving very high targeting efficiency by utilizing targeting vectors that utilize promoter-less selection cassettes and which are engineered to targeted into transcriptionally active loci. In particular, the invention provides a method for targeting promoter-less selection cassettes into transcriptionally active loci in stem cells or other eukaryotic cells with much greater efficiency than previously observed with other methods, thus reducing the number of drug-resistant clones to be screened or eliminating the need to screen for targeted cells altogether. The invention also encompasses the DNA targeting vectors, the targeted cells, as well as non-human organisms, especially mice, created from the targeted cells.

Abstract: This is intended to provide a technique for providing a stem cell having a mutation in both alleles (a pair of alleles). A method for producing a stem cell having a mutation in both chains of alleles which comprises: A) the step of providing a stem cell; B) the step of preventing Blm alleles from functioning in the stem cell; and C) the step of inducing mutation in the stem cell. It is also intended to provide a library of stem cells having a mutation in both chains of alleles wherein stem cells involved in the library have the mutation transferred thereinto over the entire genome.

Abstract: Methods are described for the delivery of one or more small interfering RNAs (siRNAs) to a eukaryotic cell using a bacterium or BTP. Methods are also described for using this bacterium to regulate gene expression in eukaryotic cells using RNA interference, and methods for treating viral diseases and disorders. The bacterium or BTP includes one or more siRNAs or one or more DNA molecules encoding one or more siRNAs. Vectors are also described for use with the bacteria of the invention for causing RNA interference in eukaryotic cells.

Abstract: Disclosed herein are methods and compositions for targeted integration of an exogenous sequence into the human PPP1R12C locus, for example, for expression of a polypeptide of interest.

Abstract: The present invention relates to double-stranded oligonucleotides, pharmaceutical compositions thereof, and use of such double-stranded oligonucleotides and pharmaceutical compositions to modulate nociceptive signaling in a cell or prevent and/or treat pain in a patient.

Abstract: A method of preparing gene trapping libraries, and gene targeted cells for conditional inactivation of genes, is provided. The invention provides a vector having a mutational element cassette and a gene trap cassette, each cassette having site-specific recombination sequences.

Abstract: Regulatory elements, specifically replicators and transgene constructs containing replicator nucleic acid sequences, are disclosed herein. Methods of using replicators and transgene constructs including replicators to inhibit, delay, or prevent gene silencing are also disclosed herein.

Abstract: The present invention relates to germ line and somatic cells comprising a mutant p27kip1 protein lacking a Cdk2 phosphorylation site. Also provided are transgenic animals and methods of making such transgenic animals which have increased size and/or growth rate.

Abstract: A process is provided of introducing an RNA into a living cell to inhibit gene expression of a target gene in that cell. The process may be practiced ex vivo or in vivo. The RNA has a region with double-stranded structure. Inhibition is sequence-specific in that the nucleotide sequences of the duplex region of the RNA and of a portion of the target gene are identical. The present invention is distinguished from prior art interference in gene expression by antisense or triple-strand methods.

Abstract: The present invention relates to the specific inhibition of expression of a target gene in mammals using a short double stranded RNA. The dsRNA is less than 49 nucleotides in length and has a nucleotide sequence which is complementary to at least a part of the target gene. The dsRNAs of the present invention are useful for treating diseases, for example, cancer, viral diseases or neurodegenerative diseases.

Abstract: The present application relates to methods and compositions for triggering RNAi. Triggered RNAi is highly versatile because the silencing targets are independent of the detection targets. In some embodiments, a method of silencing a target gene is provided.

Abstract: The disclosure relates generally to stem cell biology and more specifically to genetic manipulation of stem cells. Methods and compositions using recombinational cloning techniques are disclosed which allow the construction and insertion of complex genetic constructs into embryonic and adult stem cells and progenitor cells. The methods disclosed will allow the harvesting of adult stem cells pre-engineered with integration sites to facilitate early passage genetic modification.

Abstract: The invention relates to methods for directing integration of a nucleic acid of interest towards homologous recombination and uses thereof. The present invention discloses factors involved in integration of a nucleic acid by illegitimate recombination which provides a method of directing integration of a nucleic acid of interest to a predetermined site, whereby the nucleic acid has a homology at or around the predetermined site, in a eukaryote with a preference for non-homologous recombination comprising steering an integration pathway towards homologous recombination. Furthermore, the invention provides a method of directing integration of a nucleic acid of interest to a subtelomeric and/or telomeric region in a eukaryote with a preference for non-homologous recombination.

Abstract: The present invention relates to transfected primary and secondary somatic cells of vertebrate origin, particularly mammalian origin, transfected with exogenous genetic material (DNA) which encodes erythropoietin or an insulinotropin [e.g., derivatives of glucagon-like peptide 1 (GLP-1)], methods by which primary and secondary cells are transfected to include exogenous genetic material encoding erythropoietin or an insulinotropin, methods of producing clonal cell strains or heterogenous cell strains which express erythropoietin or an insulinotropin, methods of gene therapy in which the transfected primary or secondary cells are used, and methods of producing antibodies using the transfected primary or secondary cells.

Abstract: Disclosed are vectors, kits and methods useful in the construction of recombinant cells and DNAs via enhanced efficiency homologous recombination. The vectors are targeting vectors that contain a gene-of-interest spliced between two ends that are homologous to a genome target site. The ends of the vector may be protected from exonuclease attack by deploying a cap, such as a hair pin structure. The vector is linked to a nuclear localization signal sequence, and preferably, a bait peptide that binds to RAD51, to facilitate homologous recombination. The vector may be deployed in myriad genetic transformation applications, such as site-directed mutagenesis, gene therapy, and the like.

Abstract: The present invention provides methods for generating and characterizing gene targeting events by using tags. More specifically, the invention employs methods to enrich for cells that have undergone the desired targeting event.

Abstract: Disclosed herein are methods and compositions for inactivation of the human glucocorticoid receptor (GR) gene by targeted cleavage of genomic DNA encoding the GR. Such methods and compositions are useful, for example, in therapeutic applications which require retention of immune function during glucocorticoid treatment.

Abstract: DNA gene inactivation constructs for homologous recombination in the genome of a mammalian cell are provided.

Abstract: The present invention relates to a mutated gene coding for a mutant LAT protein leading to exaggerated TH2 differentiation. The invention relates to biological structures containing the mutant, particularly non-human LAT gene, mutated animals, plasmids, chromosomal DNAs, embryos comprising the mutated gene, and applications thereof. The invention further relates to screening methods for drugs useful for treatment against asthma and allergy. Otherwise, the invention relates to methods for producing IgE antibodies.

Abstract: The present invention provides nucleic acids that include a nucleotide sequence that encodes an siRNA, which nucleotide sequence is operably linked to a target cell-specific promoter RNA polymerase II promoter. The present invention further provides vectors, including expression vectors, which include a subject nucleic acid; and host cells that harbor a subject nucleic acid or a subject expression vector. The present invention further provides methods of modulating (e.g., reducing) expression of a gene in a target cell-specific manner, the methods generally involving introducing into a cell a subject expression vector.

Abstract: The present invention relates to a Drosophila in vitro system which was used to demonstrate that dsRNA is processed to RNA segments 21-23 nucleotides (nt) in length. Furthermore, when these 21-23 nt fragments are purified and added back to Drosophila extracts, they mediate RNA interference in the absence of long dsRNA. Thus, these 21-23 nt fragments are the sequence-specific mediators of RNA degradation. A molecular signal, which may be their specific length, must be present in these 21-23 nt fragments to recruit cellular factors involved in RNAi. This present invention encompasses these 21-23 nt fragments and their use for specifically inactivating gene function. The use of these fragments (or chemically synthesized oligonucleotides of the same or similar nature) enables the targeting of specific mRNAs for degradation in mammalian cells, where the use of long dsRNAs to elicit RNAi is usually not practical, presumably because of the deleterious effects of the interferon response.

Abstract: The invention relates to DNA-sequences, especially transcription- or expression-enhancing elements (TE elements) and their use on an expression vector in conjunction with an enhancer, a promoter, a product gene and a selectable marker. The invention describes Sequence No. 1 and TE elements TE-01, -02, -03, -04, -06, -07, -08, -10, -11 or -12. Because of their small size, TE-06, TE-07 or TE-08 are particularly preferred. Sequence No. 1 originates from a sequence region located upstream from the coding region of the Ub/S27a gene from CHO cells. TE elements bring about an increase in the expression of the product gene, particularly when stably integrated in the eukaryotic genome, preferably the CHO-DG44 genome. Chromosomal positional effects are thereby overcome, shielded or cancelled out. In this way the proportion of high producers in a transfection mixture and also the absolute expression level are increased up to seven-fold.

Abstract: The present invention provides an improved method for achieving efficient transcription and translation of modified transgene constructs in vector systems. The vector may be a lentiviral vector. Such a method facilitates the production of viral vector genomes with intact functional transgene sequences allowing stable integration of a transgene-containing viral vector genome into the germline of an animal such as a transgenic avian. The subsequent expression of the transgene results in a recombinant protein product being produced, which, in the case of a transgenic avian can result in the targeted production of the protein into the egg of the transgenic bird.

Abstract: The invention relates to an in vitro method for introducing a targeted genome modification into an oocyte or an egg and a method for performing a random insertion in the genome of a host cell.

Abstract: Mutational inactivation of proteins involved in para-cresol production in certain milk products results in improved taste and odor. The undesirable para-cresol forms over time as a result of enzymes produced by the bacterium that produces gellan gum. Since the gellan is typically used in a relatively unpurified form, the enzymes are added to the milk along with the gellan. Inactivation of the enzymes is a genetic means of eliminating the enzymes without requiring any additional purification or processing.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: Detection of phenols using engineered bacteria. A biosensor can be created by placing a reporter gene under control of an inducible promoter. The reporter gene produces a signal when a cognate transcriptional activator senses the inducing chemical. Creation of bacterial biosensors is currently restricted by limited knowledge of the genetic systems of bacteria that catabolize xenobiotics. By using mutagenic PCR to change the chemical specificity of the Pseudomonas species CF600 DmpR protein, the potential for engineering novel biosensors for detection of phenols has been demonstrated. DmpR, a well-characterized transcriptional activator of the P. CF600's dmp operon mediates growth on simple phenols. Transcription from Po, the promoter heading the dmp operon, is activated when the sensor domain of DmpR interacts with phenol and mono-substituted phenols.

Abstract: The invention relates to Infectious Bursal Disease Virus (“IBDV”) and vaccines therefor. Provided are infectious recombinant Infectious Bursal Disease Virus (“rIBDV”) essentially incapable of growing in a cell that is not derived from a bursa cell, or an infectious rIBDV having retained at least part of the very virulent characteristics of a very virulent Infectious Bursal Disease Virus (“vvIBDV”).

Abstract: This invention relates to methods and compositions for obtaining Arabidopsis and Brassica plants. Specifically, the method provides culturing protocols and compositions that facilitate the regeneration of transformed plants following delivery of beneficial DNA molecules.

Abstract: The Invention pertains to methods, kits, molecules and cells to increase the rate or recombination and/or target recombination in dividing cells. In a particular aspect, the invention concerns methods and kits to induce targeted meiotic recombination.

Abstract: The invention pertains to methods, kits, molecules and cells to increase the rate or recombination and/or target recombination in dividing cells. In a particular aspect, the invention concerns methods and kits to induce targeted meiotic recombination.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: A packaging cell line capable of complementing recombinant adenoviruses based on serotypes from subgroup B, preferably adenovirus type 35. The cell line is preferably derived from primary diploid human cells transformed by adenovirus E1 sequences either operatively linked on one or two DNA molecules, the sequences operatively linked to regulatory sequences enabling transcription and translation of encoded proteins. Also, a cell line derived from PER.C6 that expresses functional Ad35-E1B sequences. The Ad35-E1B sequences are driven by the E1B promoter and terminated by a heterologous poly-adenylation signal. The new cell lines are useful for producing recombinant adenoviruses. The cell lines can be used to produce human recombinant therapeutic proteins such as human antibodies. In addition, the cell lines are useful for producing human viruses other than adenovirus such as influenza, herpes simplex, rotavirus, and measles.

Abstract: Disclosed are isolated transposable elements, or isolated DNA sequences which encode a transposase protein or a portion of a transposase protein. The isolated transposable elements or the isolated DNA sequences are members of the mPing/Pong family of transposable elements. The invention also relates to a purified transposase protein, or peptide fragments thereof, encoded by such DNA sequences. Such transposable elements are useful in applications such as the stable introduction of a DNA sequence of interest into a eukaryotic cell. The sequence information disclosed herein is useful in the design of oligonucleotide primers which are useful for the isolation of related members of the mPing/Pong family of transposable elements, or for the detection of transpositions of the transposable elements.

Abstract: Presented are methods and compositions for targeted chromosomal genomic alterations using modified single-stranded oligonucleotides of the invention have at least one modified nuclease-resistant terminal region comprising phosphorothioate linkages, LNA analogs or 2?-O-Me base analogs.

Abstract: Cleavage site for the protease furin is inserted between domains of a membrane glycoprotein. Upon cleavage by furin in the trans-Golgi network, the protein is separated into individual membrane-free domain that retains its native conformation. This protocol can be used to produce virus membrane protein domains for structural analysis and for trials as vaccines.

Abstract: Methods are provided for isolation of DNA sequences encoding proteins with properties of interest by means of expression cloning in filamentous fungal host cells. The isolated DNA sequences are useful in processes for producing the proteins of interest.

Abstract: An isolated DNA encoding the enzyme I-SceI is provided. The DNA sequence can be incorporated in cloning and expression vectors, transformed cell lines and transgenic animals. The vectors are useful in gene mapping and site-directed insertion of genes.

Abstract: The invention concerns human cells which, due to an activation of the endogenous human EPO gene, are able to produce EPO in an adequate quantity and purity to enable a cost-effective production of human EPO as a pharmaceutical preparation. Furthermore the invention concerns a process for the production of such human EPO-producing cells, DNA constructs for the activation of the endogenous EPO gene in human cells as well as a process for the large-scale production of EPO in human cells.