Abstract: Described herein are immunodeficient non-human animals lacking expression of toll-like receptor 4 (TLR4) by endogenous autogeneic innate immune cells, as well as methods and compositions for engraftment of xenogeneic hematopoietic stem cells in the immunodeficient non-human animal lacking expression of toll-like receptor 4 (TLR4), thereby creating an innate immune system in the animal derived from the xenogeneic hematopoietic stem cells. Further described are immunodeficient mice lacking expression of toll-like receptor 4 by endogenous autogeneic innate immune cells, as well as methods and compositions for engraftment of xenogeneic hematopoietic stem cells in the immunodeficient mouse lacking expression of toll-like receptor 4, thereby creating an innate immune system in the animal derived from the xenogeneic hematopoietic stem cells.

Abstract: The invention relates to a minicircle transfer vector for producing viral vectors comprising a transfer sequence and specific packing signals flanking both sides of the transfer sequence for packaging of the transfer sequence into particles of a viral vector. The invention also relates to minicircle packaging vectors carrying support functions for producing viral vectors. The invention further relates to cells bearing the disclosed minicircles. The invention further relates to methods for producing viral vectors using such minicircles and viral vectors obtained thereby, as well as kits useful in performing the described methods.

Abstract: Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

Abstract: The present invention is within the field of industrial protein production. The inventors have designed and constructed a new expression system comprising an expression vector coding for a glutamine synthetase of human or dog origin, and a CHO cell line. More specifically, the invention pertains to a combination of (i) a DNA vector suitable for production of a recombinant protein, wherein said vector comprises a sequence coding for a glutamine synthetase, and (ii) a Chinese Hamster Ovary (CHO) cell line, wherein said GS comprises a sequence at least 94.5% identical to the sequence of SEQ ID NO: 1 or to the sequence of SEQ ID NO: 2.

Abstract: A mutant Pichia pastoris alcohol oxidase 1 (AOX1) promoter of the wild type Pichia pastoris AOX1 promoter (SEQ ID No. 1) comprising at least one mutation selected from the group consisting of: a) a transcription factor binding site (TFBS), b) nucleotides 170 to 235 (?784 to ?719), nucleotides 170 to 191 (?784 to ?763), nucleotides 192 to 213 (?762 to ?741), nucleotides 192 to 210 (?762 to ?744), nucleotides 207 to 209 (?747 to ?745), nucleotides 214 to 235 (?740 to ?719), nucleotides 304 to 350 (?650 to ?604), nucleotides 364 to 393 (?590 to ?561), nucleotides 434 to 508 (?520 to ?446), nucleotides 509 to 551 (?445 to ?403), nucleotides 552 to 560 (?402 to ?394), nucleotides 585 to 617 (?369 to ?337), nucleotides 621 to 660 (?333 to ?294), nucleotides 625 to 683 (?329 to ?271), nucleotides 736 to 741 (?218 to ?213), nucleotides 737 to 738 (?217 to ?216), nucleotides 726 to 755 (?228 to ?199), nucleotides 784 to 800 (?170 to ?154) or nucleotides 823 to 861 (?131 to ?93) of Seq ID No.

Abstract: Subjecting a heterogeneous cell population (one with both stem cells and non-stem cells) to extreme stress selectively eliminated the non-stem cells and resulted in the enrichment of stem cells in the population. The stress can take many forms, including without limitation, cell toxins, high temperature, high salt, and low oxygen (hypoxic) conditions. The number of stem cells remaining after stress were increased, and showed increased expression of traditional stem cell markers. The stem cells were shown to be capable of proliferation and differentiation into multiple types of cells. This method allows purification of stem cells from adult heterogeneous cell populations on a large scale basis without requirement of expensive equipment, and without requiring the presence of cell surface markers. Stem cells produced by the above method can be used for clinical applications, including tissue engineering.

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: Methods are described that bias cells, such as potent and multipotent stem cells, by transfection with a nucleic acid sequence, to differentiate to a desired end-stage cell or a cell having characteristics of a desired end-stage cell. In particular embodiments, human neural stem cells or mesenchymal stem cells are transfected with vectors comprising genes in the homeobox family of transcription factor developmental control genes, and this results in a greater percentage of resultant transformed cells, or their progeny, differentiating into a desired end-stage cell or a cell having characteristics of a desired end-stage cell.

Abstract: Efficient and muscle-specific gene expression can be obtained with constructs containing two or more copies of USE and/or ?USE fused to the minimal promoter of the TnISlow gene. USE is a small (about 160-bp) upstream enhancer of the TnISlow gene that confers slow-twitch muscle fiber specificity. ?USE is generated from a 100-bp deletion at the 5? end of USE. ?USE confers expression in slow- and fast-twitch muscle fibers. The strength and relatively small size (less than 600-bp) of these constructs make them useful for gene therapy applications.

Abstract: This invention relates to industrial production of proteins. More specifically, the invention relates to the res-DHFR surrogate marker, which corresponds to a fusion between DHFR and a protein conferring resistance to a toxic compound or conferring a metabolic advantage. The invention further relates to the use of res-DHFR for screening cells for high expression of a protein of interest. The invention is illustrated by the Puro-DHFR surrogate marker, which corresponds to a fusion between the puromycin N-acetyltransferase and dihydrofolate reductase (DHFR).

Abstract: The present invention refers to a cell culture system especially for investigating the sensitizing, allergenic and/or irritating effect of substances, comprising a first and a second compartment that can communicate with each other via a permeable interlayer, whereby the first compartment has an epidermis model and the second a cell culture based on immune cells.

Abstract: The present invention encompasses an expression vector that is capable of generating a virus from a segmented genome. In particular, a single expression vector may be utilized to produce influenza virus in cultured cells. The expression vector can be delivered in a purified DNA form or by a suitably designed bacterial carrier to cells in culture or to animals. This invention increases the virus generation efficiency, which benefits vaccine development. The bacterial carrier harboring such a plasmid encoding an attenuated virus may be used as a vaccine against corresponding viral disease.

Abstract: The present invention provides novel compositions to transfect cells for production of growth hormone (GH). These novel compositions also are used to produce germline transgenic birds that can successfully pass the transgene encoding growth hormone to their offspring. These novel compositions include components of vectors such as a vector backbone, a novel promoter, and a gene of interest that encodes for GH, and the vectors comprising these components. In one embodiment these vectors are transposon-based vectors. The present invention also provides methods of making these compositions and methods of using these compositions for the production of GH in vitro and in vivo. In one embodiment the GH is human (h)GH.

Abstract: Novel compositions for the production of the light and heavy chains of an antibody are provided. These novel compositions are also used to produce germline transgenic birds that can successfully pass the transgene encoding the antibody to their offspring. The compositions comprise components of vectors, such as a vector backbone, a promoter, and a gene of interest that encodes the light or heavy chain of an antibody, and the vectors comprising these components. In certain embodiments, these vectors are transposon-based vectors. Also provided are methods of making these compositions and methods of using these compositions for the production of the light and heavy chains of an antibody. In one embodiment, the antibody is a human monoclonal antibody.

Abstract: Novel compositions for the in vitro or in vivo production of specific proteins are provided. The compositions comprise components of vectors, such as a vector backbone, a promoter, and a gene of interest that encodes for the protein of interest, and the transposon-based vectors comprising these components. Also provided are methods of making these compositions and methods of using these compositions for the production of desired proteins in vivo or in transfected cells in vitro.

Abstract: Compositions and methods are provided for the rapid and highly accurate identification of the entire essential genome of any organism under a given selection condition at a resolution of a few base pairs. An engineered transposon bearing an adapter sequence for ultra high throughput adaptor-based sequencing is employed for hyper-saturated transposon mutagenesis. Transposon junctions are subsequently isolated and collectively amplified through a shared parallel PCR strategy such that a second adaptor sequence is further incorporated into template DNA so that the first adaptor sequence and the second adaptor sequence flank the 5? and 3? regions of the sample DNA, respectively. Sample DNA is then sequenced in an ultra high-throughput adaptor-based DNA sequencer using adaptor primers. Transposon insertion sites are mapped onto the organism's genome, allowing for the algorithmic identification of essential genetic elements based on genomic transposition frequency.

Abstract: This invention provides compositions and methods for producing translational components that expand the number of genetically encoded amino acids in vertebrate cells. The components include orthogonal tRNA's, orthogonal aminoacyl-tRNA synthetases, orthogonal pairs of tRNA's/synthetases and unnatural amino acids. Proteins and methods of producing proteins with unnatural amino acids in vertebrate cells are also provided. The present invention provides vertebrate cells with translation components, e.g., pairs of orthogonal aminoacyl-tRNA synthetases (O-RSs) and orthogonal tRNA's (O-tRNA's) and individual components thereof, that are used in vertebrate protein biosynthetic machinery to incorporate an unnatural amino acid in a growing polypeptide chain, in a vertebrate cell.

Abstract: An object of the present invention is to provide a screening system that targets ion channels and has superior efficiency. The present invention provides a material for screening for compounds that act on a target ion channel, comprising cells which retain at least one first DNA encoding a voltage-dependent Na ion channel that has been inhibited from being inactivated, and in which a K ion channel has been activated so that a resting membrane potential becomes deeper in a negative direction.

Abstract: The present invention provides a pseudo-tissue for accuracy control comprising a nucleic acid or cells, a holding body for holding the nucleic acid or cells, and a protecting body for covering at least a part of the surface of the holding body so as to protect the holding body. A method for controlling accuracy by using the pseudo-tissue, and a method for manufacturing the pseudo-tissue are also disclosed.

Abstract: This invention relates to a lamellae tissue layer, comprising a grooved silk fibroin substrate comprising tissue-specific cells. The silk fibroin substrates provides an excellent means of controlling and culturing cell and extracellular matrix development. A multitude of lamellae tissue layers can be used to create a tissue-engineered organ, such as a tissue-engineered cornea. The tissue-engineered organ is non-immunogenic and biocompatible.