Abstract: Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3?-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.

Abstract: Methods and perfusion culture systems are disclosed. The systems and methods relate to decreasing the starting perfusion rate, resulting in increased residence time of the cells in the bioreactor and the cell retention device, and/or concomitantly increasing the starting bioreactor volume or decreasing the starting cell retention device volume, or both. Other method embodiments include increasing the concentrations of individual components of the tissue culture fluid, and adding a stabilizer of the degradation of the recombinant protein.

Abstract: The present invention is directed to a method of deriving pluripotent embryonic stem cells from mouse blastocysts or from primordial germ cells from a post-implantation mouse embryo, or of maintaining or growing pluripotent embryonic stem cells from a mouse, or of expanding human hematopoietic stem cells or human hematopoietic precursor cells. The methods include the step of cultivating the stem cells or precursor cells for at least one passage in a culture medium preconditioned by the rabbit fibroblast cell line Rab9 (ATCC catalogue CRL1414) and containing less than 0.1 ng/ml Leukemia Inhibitory Factor (LIF).

Abstract: Disclosed herein are isolated immunogens including variant gp120 polypeptides. In an example, a variant gp120 polypeptide includes a deletion of at least 8 consecutive residues of the fourth conserved loop (C4) between residues 419 and 434 of gp120 according to HXB2 numbering. Also provided are isolated nucleic acid molecules encoding the disclosed isolated immunogens. In an example, an isolated nucleic acid molecule further includes a nucleic acid molecule encoding a hepatitis B surface antigen or a variant thereof. Compositions including the isolated immunogens including variant gp120 polypeptides are also disclosed. In some examples, a composition further includes a carrier protein, such as a hepatitis B surface antigen or a variant thereof (natural or recombinant). Viral-like particles are also provided including any of the disclosed isolated immunogens or compositions.

Abstract: The invention relates to conjugates comprising a targeting moiety specific for the CXCR4 based on the polyphemusin-derived peptide and a therapeutic or imaging agent. The invention relates as well to the application of said conjugates for the therapy and diagnostics which require the specific targeting to CXCR4+ cells.

Abstract: A method of deriving isolated stem cells including: implanting a matrix in a wound site of a living organism; allowing cells to infiltrate the matrix; removing the matrix containing the infiltrated cells from the wound site; and removing the infiltrated cells from the matrix to provide isolated stem cells. Stem cells produced by this process, stem cells with certain characteristics, and methods for treating wounds using these stem cells are provided.

Abstract: The invention provides methods for detecting the presence of cancer initiating cells in a tissue, methods for identifying test agent for reducing cancer, and methods for reducing cancer in a subject. The invention's methods are applicable to any cancer, and in particular to liver cancer. The invention also provides the isolation and characterization of pre-malignant hepatocellular carcinoma initiating cells (HICs). The invention further provides methods for isolating hepatocellular carcinoma initiating cells (HICs), methods for using the isolated cells for screening anti-cancer drugs, methods for using HIC markers for the early diagnosis of hepatocellular carcinoma, and methods for the prevention and/or delay of hepatocellular carcinoma by using agents that selectively deplete the number and/or malignant properties of HICs.

Abstract: Described are compounds and methods useful in the promotion of muscle growth, the treatment of muscle loss or insufficient muscle growth, and the treatment of fibrotic conditions.

Abstract: The present invention relates to a method of collecting semen from the epididymis or the testis of a lab animal and an artificial insemination method thereof. The present invention relates to a new applicable method in the animal production field using artificial insemination. Since the present invention solves the problems of the conventional method of artificial insemination of lab animals, it is excellent in terms of the pregnancy rate and productivity; is time-efficient; and is superior in economic and industrial respects by preventing a huge economic loss due to the costs for maintaining the mass breeding of animals.

Abstract: The present invention relates to a novel protein skeletal module which increases the binding affinity or binding specificity of active polypeptides. More particularly, the present invention relates to a protein skeletal module comprising polypeptides consisting of the 1st to 19th amino acids of the amino acid sequence expressed in sequence number 1; polypeptides comprising active polypeptides; and polypeptides consisting of the 29th to 86th amino acids of the amino acid sequence expressed in sequence number 1. The present invention also relates to a method for preparing the protein skeletal module. The protein skeletal module of the present invention increases the binding affinity or binding specificity of active polypeptides embedded therein, and therefore is effective in diagnosing and treating diseases.

Abstract: The invention provides a CR2-FH molecule comprising a CR2 portion comprising CR2 protein or a fragment thereof and a FH portion comprising a factor H protein or a fragment thereof, and pharmaceutical compositions comprising a CR2-FH molecule. Also provided are methods of using the compositions for treatment diseases in which the alternative complement pathway is implicated, such as age-related macular degeneration, rheumatoid arthritis, and ischemia reperfusion.

Abstract: The present invention relates to modified nucleic acid sequences coding for coagulation factor VIII (FVIII) and for von Willebrand factor (VWF) as well as complexes thereof and their derivatives, recombinant expression vectors containing such nucleic acid sequences, host cells transformed with such recombinant expression vectors, recombinant polypeptides and derivatives coded for by said nucleic acid sequences which recombinant polypeptides and derivatives do have biological activities together with prolonged in vivo half-life and/or improved in vivo recovery compared to the unmodified wild-type protein. The invention also relates to corresponding FVIII sequences that result in improved expression yield. The present invention further relates to processes for the manufacture of such recombinant proteins and their derivatives. The invention also relates to a transfer vector for use in human gene therapy, which comprises such modified nucleic acid sequences.

Abstract: Non-human totipotent or pluripotent cells are provided comprising at a genomic locus a self-excisable, recombinase expression cassette flanked with recombination recognition sites, wherein a recombinase gene is operably linked to a promoter that is active in a post-meiotic spermatid stage when cytoplasmic bridging occurs between spermatids. Compositions and methods are provided for making cassette-deleted F1 non-human animals, wherein the methods comprise employing totipotent or pluripotent cells containing a self-excisable, recombinase expression cassette.

Abstract: This invention relates to the field of biotechnology or genetic engineering. Specifically, this invention relates to the field of gene expression. More specifically, this invention relates to novel substitution mutant receptors and their use in a Group H nuclear receptor-based inducible gene expression system and methods of modulating the expression of a gene in a host cell for applications such as gene therapy, large scale production of proteins and antibodies, cell-based high throughput screening assays, functional genomics and regulation of traits in transgenic organisms.

Abstract: This disclosure relates to vectors, isolated cells, compositions, and methods for the treatment of critical limb ischemia and associated disorders. One aspect of the disclosure relates to a vector comprising a nucleic acid encoding a 165A isoform VEGF protein and a promoter that regulates expression of the nucleic acid encoding the VEGF.

Abstract: The present disclosure provides a DNA-targeting RNA that comprises a targeting sequence and, together with a modifying polypeptide, provides for site-specific modification of a target DNA and/or a polypeptide associated with the target DNA. The present disclosure further provides site-specific modifying polypeptides. The present disclosure further provides methods of site-specific modification of a target DNA and/or a polypeptide associated with the target DNA The present disclosure provides methods of modulating transcription of a target nucleic acid in a target cell, generally involving contacting the target nucleic acid with an enzymatically inactive Cas9 polypeptide and a DNA-targeting RNA. Kits and compositions for carrying out the methods are also provided. The present disclosure provides genetically modified cells that produce Cas9; and Cas9 transgenic non-human multicellular organisms.

Abstract: The instant invention relates to the use of 24-hydroxylated vitamin D compounds as therapeutics in mammalian bone fracture repair. In addition, the instant invention relates to novel 24-hydroxylated vitamin D compound receptors which can be employed in the development of compounds capable of facilitating fracture repair in animals. The instant invention also relates to nucleic acids encoding such receptors as well as vectors, host cells, transgenic animals comprising such nucleic acids and screening assays employing such receptors.

Abstract: Embodiments are directed compositions related to Eilat virus and uses thereof.

Abstract: The present invention is directed generally to cell culture media useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells in the presence of said media. Cells containing introduced materials can be further cultured in the media. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells). The invention thus provides efficient and high throughput methods to transform/transfect culture and cells avoiding the need for multiple manipulations and transfers of cells during transfection and expression studies.

Abstract: The invention relates to mammalian haploid embryonic stem cells and methods for the production of such stem cells. The inventions also relates to a cell culture and a cell line of mammalian haploid embryonic stem cells.

Abstract: The instant invention relates to the use of 24-hydroxylated vitamin D compounds as therapeutics in mammalian bone fracture repair. In addition, the instant invention relates to novel 24-hydroxylated vitamin D compound receptors which can be employed in the development of compounds capable of facilitating fracture repair in animals. The instant invention also relates to nucleic acids encoding such receptors as well as vectors, host cells, transgenic animals comprising such nucleic acids and screening assays employing such receptors.

Abstract: This invention relates to double-stranded ribonucleic acid (dsRNA), and its use in mediating RNA interference to inhibit the expression of an RNA from the West Nile virus (WNV), and the use of the dsRNA to treat pathological processes mediated by WNV infection, such as viral encephalitis.

Abstract: The present invention relates to a model animal spontaneously developing anemia. More specifically, the invention relates to a transgenic non-human mammal spontaneously developing anemia associated with a postnatal decrease in production of erythropoietin (Epo), Epo-producing cells prepared from the transgenic non-human mammal, and a screening method using the Epo-producing cells.

Abstract: The present disclosure relates to methods of decreasing lactate production in cell culture using divalent transitional metallic salts. The present disclosure also relates to a method of producing polypeptide by adding divalent transitional metallic salt to the cell culture medium for reducing lactate accumulation followed by fermenting and recovering the polypeptide.

Abstract: The present invention relates a combination for use in the treatment and/or prevention of mastitis containing i) an agonistic anti-CD40 monoclonal antibody or a CD40 ligand or a vector coding for the anti-CD40 antibody or a vector coding for the CD40L; and ii) inactivated or attenuated bacteria selected from the group consisting of Staphylococcus, Streptococcus, Listeria or Escherichia.

Abstract: The present invention provides a pharmaceutical composition comprising a protein having ?-galactosidase activity for treating Fabry disease, which causes no allergic side effect, which is highly stable in blood (plasma) and which can readily be taken up by a cell of an affected organ. The pharmaceutical composition for treating Fabry disease of the invention comprises, for example, a protein which acquires an ?-galactosidase activity through alteration of the structure of the active site of wild-type human ?-N-acetylgalactosaminidase.

Abstract: The present invention concerns constructs based on sequences derived from the partitioning system of plasmid and chromosomal DNA of bacteria, such as eukaryotic expression vectors, fusion proteins and polynucleotides encoding the same and also eukaryotic cells transformed with or expressing such constructs. The present invention also concerns the use thereof in the regulation of gene expression and/or in the detection and control of the dynamics, localization or metabolism of genomic DNA loci of interest in eukaryotic cells.

Abstract: The present invention relates antidotes to anticoagulants targeting factor Xa. The antidotes are factor Xa protein derivatives that bind to the factor Xa inhibitors thereby substantially neutralizing them but do not assemble into the prothrombinase complex. The derivatives describe herein lack or have reduced intrinsic coagulant activity. Disclosed herein are methods of stopping or preventing bleeding in a patient that is currently undergoing anticoagulant therapy with a factor Xa inhibitor.

Abstract: There is provided an expression vector for animal cell having an increased gene expression efficiency, and particularly, an expression vector for animal cells including a MAR element and a SAR element, which are gene expression increasing factors, at a 5? end of a promoter, a 3? end of a transcription termination site, or at both of the 5? end of the promoter and the 3? end of the transcription termination site. The expression vector for animal cells according to the present invention exhibits remarkably increased gene expression efficiency as compared to conventional expression vectors for animal cells, such that protein expression of foreign genes may be significantly increased using this expression vector for animal cells. Particularly, the expression vector for animal cells according to the present invention may be useful in that a high-expression cell line may be secured even without MTX amplification.

Abstract: Disclosed are compositions and methods for increasing the longevity of a cell culture and permitting the increased production of proteins, preferably recombinant proteins, such as antibodies, peptides, enzymes, growth factors, interleukins, interferons, hormones, and vaccines. Cells transfected with an apoptosis-inhibiting gene or vector, such as a triple mutant Bcl-2 gene, can survive longer in culture, resulting in extension of the state and yield of protein biosynthesis. Such transfected cells exhibit maximal cell densities that equal or exceed the maximal density achieved by the parent cell lines. Transfected cells can also be pre-adapted for growth in serum-free medium, greatly decreasing the time required to obtain protein production in serum-free medium. In certain methods, the pre-adapted cells can be used for protein production following transformation under serum-free conditions. The method preferably involves eukaryotic cells, more preferably mammalian cells.

Abstract: Provided is a polynucleotide encoding a protein having an activity to transfer a sugar to the hydroxy groups at the 4?- and 7-positions of a flavone.

Abstract: Methods for producing compositions of decellularized extracellular matrix (DM) tissue culture are described. The compositions can be used for coating supports such as tissue culture substrates, osteogenic gels, and medical devices.

Abstract: Pharmaceutical compositions and methods for inhibiting cell growth, modulating function of PCNA, treating prostate cancer, and enhancing PCNA trimer formation are disclosed.

Abstract: The present invention relates to a method for the production of differentiated respiratory epithelial cells comprising: (a) providing a cell population comprising or consisting of precursor cells of respiratory epithelial cells; (b) culturing the cell population of (a) in culture medium to which keratinocyte growth factor has been added; wherein the cultured cell population is supplemented with a glucocorticoid, a cAMP analogue and a cAMP elevating agent and wherein said supplementation is either simultaneously with the addition of keratinocyte growth factor in step (b) or prior or subsequently to the addition of keratinocyte growth factor in step (b), thereby differentiating said precursor cells into respiratory epithelial cells.

Abstract: The present invention relates to immunogenic peptides and their various applications. In particular the invention relates to immunogenic peptides derived from the PASD1 protein and their use in therapeutic, diagnostic and prognostic methods.

Abstract: The invention relates to a set of genetic constructs which allow the efficient and reproducible introduction of a specific nucleotide sequence at a fixed position in the genome by generating a double strand break at a specific position in the genome using a meganuclease and so stimulating a homologous recombination event at this locus between the genomic site and a transfected donor sequence. The present invention also relates to methods using these constructs and to these materials in the form of a kit.

Abstract: A micro fluidic device comprises one microstructure layer (5) and one cover layer (1), wherein the cover layer (1) is connected to the microstructure layer (5). The microstructure layer (5) comprises one bottom layer and a plurality of microstructures on it to position samples. The cover layer (1) comprises one top layer, one positioning well (6) and at least one inlet pool (4). The positioning well (6) is right above the microstructures and connected with each other. The inlet pools (4) and the positioning well (6) are connected by microchannels (3) which are formed between the microstructure layer (5) and the cover layer (1). The micro fluidic device can be applied in vitro fertilization, in determining how glial cells affect neurons, in constructing neural network and in detecting cell growth conditions.

Abstract: The present invention relates to an expression cassette useful for the expression of a polynucleotide sequence encoding a polypeptide.

Abstract: The invention provides human cells, particularly human T cells, comprising a murine T Cell Receptor (TCR) having antigen specificity for the cancer antigen gp100. Isolated or purified TCRs having antigenic specificity for amino acids 154-162 of gp100 (SEQ ID NO: 1), as well as related polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, populations of cells, antibodies, or antigen binding fragments thereof, conjugates, and pharmaceutical compositions, are further provided. The invention further provides a method of detecting the presence of cancer in a host and a method of treating or preventing cancer in a host comprising the use of the inventive materials described herein.

Abstract: The present invention provides compositions and methods relating to IL-1Rrp2 requiring proteins.

Abstract: Two vIRF4 (Kaposi's-sarcoma-associated-herpesvirus vIRF4) peptides, vif1, corresponding to aa202-216 of vIRF4, and vif2, corresponding to aa220-236 of vIRF4, are potent and selective HAUSP antagonists. The vif1 and vif2 peptides robustly suppress HAUSP DUB enzymatic activity, ultimately leading to p53-mediated anti-cancer activity. The vif1 and vif2 peptides, along with their homologues, are useful in treating cancer through regulation of p53 activity in a cancer cell. Also disclosed is the crystalline structure of vIRF4-HAUSP TRAF domain complex. The structure is useful in computer aided drug design for identifying an agent that interacts with and inhibits HAUSP, resulting in p53 medicated cell cycle arrest of cancer cells.

Abstract: Mice, embryos, cells, and tissues having a restricted immunoglobulin heavy chain locus and an ectopic sequence encoding one or more ADAM6 proteins are provided. In various embodiments, mice are described that have humanized endogenous immunoglobulin heavy chain loci and are capable of expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof that is functional in a male mouse. Mice, embryos, cells, and tissues having an immunoglobulin heavy chain locus characterized by a single human VH gene segment, a plurality of human DH gene segments and a plurality of human JH gene segments and capable expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof are also provided.

Abstract: The present application relates to a system for designing promoters for selective expression of genes. Thereby identified transcription regulatory elements are selected according to a specific methodology and used to create a library of transcription regulatory elements, which are then used to construct specific promoters, especially tissue-specific promoters.

Abstract: The present invention relates to a non-human animal deficient in the N-terminal domain of the IL-33 gene. Also provided herein is the use of said non-human animal as an in vivo model of inflammatory diseases, especially with regard to screening methods for anti-inflammatory compounds, and methods for evaluating and optimising the pharmacological properties of a given anti-inflammatory compound.

Abstract: A method, called GETWISE, for targeting mouse genes is described. GETWISE is designed to increase the frequency of homologous recombination, facilitate screening, widen the applicability of engineered animals and circumvent intrinsic gene targeting problems. GETWISE utilizes the principle of modulating gene expression by targeting tetracycline-responsive elements into a specific locus. In GETWISE alleles, control of gene expression is transferred from the endogenous to a tetracycline-inducible promoter. Endogenous promoters now control expression of the reporter gene luciferase. Breeding of GETWISE carriers with tTA/rtTA carriers enables investigators to modulate gene expression in a ubiquitous or tissue-specific manner, depending on the presence of doxycycline. GETWISE enables the study of loss or gain of gene expression in any tissue of choice within a single mouse strain. GETWISE enables the analysis of the gene expression pattern with the luciferase assay.

Abstract: The present invention relates to a polypeptide having interleukin-10 function, comprising two interleukin-10 monomer subunits covalently linked by a linker. The present invention further relates to a nucleic acid molecule encoding the polypeptide of the invention, a vector comprising said nucleic acid molecule, a non-human host transformed with the nucleic acid molecule or the vector of the invention as well as a method for the production of a recombinant polypeptide of the invention. The present invention further relates to a pharmaceutical composition as well as to the polypeptide, the nucleic acid molecule, the vector or the host or host cell of the invention for use in treating and/or preventing inflammatory diseases.

Abstract: Provided herein is a synthetic or isolated polynucleotide encoding a mammalian 18S rRNA that is resistant to pactamycin. The pactamycin-resistance is conferred by one or more single residue substitutions in the 18S rRNA sequence; a fragment thereof harboring said substitutions; a complementary sequence thereto; or a substantially identical sequence of the foregoing. Related systems, methods and kits are also described.

Abstract: The present invention relates to a novel enhancer of protein production in host cells. It discloses a vector for expressing recombinant proteins in these cells, comprising a nucleotide sequence encoding a) a secretion peptidic signal, b) a 6-methylguanine-DNA-methyltransferase enzyme (MGMT, EC 2.1.1.63), a mutant or a catalytic domain thereof, and c) a recombinant protein. Said MGMT enzyme is preferably the so-called SNAP protein.

Abstract: Compositions and methods are provided for making one or more targeted genetic modifications at a target genomic locus by employing homologous recombination facilitated by single or double-strand break at or near the target genomic locus. Compositions and methods for promoting efficiency of homologous recombination between an LTVEC and a target genomic locus in prokaryotic or eukaryotic cells using engineered nucleases are also provided.

Abstract: Targeting constructs and methods of using them are provided for differentiation-dependent modification of nucleic acid sequences in cells and in non-human animals. Targeting constructs comprising a promoter operably linked to a recombinase are provided, wherein the promoter drives transcription of the recombinase in an differentiated cell but not an undifferentiated cell. Promoters include Blimp1, Prm1, Gata6, Gata4, Igf2, Lhx2, Lhx5, and Pax3. Targeting constructs with a cassette flanked on both sides by recombinase sites can be removed using a recombinase gene operably linked to a 3?-UTR that comprises a recognition site for an miRNA that is transcribed in undifferentiated cells but not in differentiated cells. The constructs may be included in targeting vectors, and can be used to automatically modify or excise a selection cassette from an ES cell, a non-human embryo, or a non-human animal.