Abstract: Materials and methods are disclosed for regulation of biological events such as target gene transcription and growth, proliferation or differentiation of engineered cells.

Abstract: There is discloses a transgenic mouse over-expressing a potassium channel BEC1, which can be used as an effective tool for screening a substance for antidementia or a substance to improve learning and memory. Also disclosed are an in vivo screening method of a substance for antidementia or a substance to improve learning and memory, which uses the learning and memory abilities of said mouse as the index, and an in vivo screening of a substance for antianxiety, which uses acceleration of anxiety as the index. In addition, there is disclosed a method for producing a pharmaceutical composition for antidementia, learning and memory improvement use or antianxiety, using a substance capable of inhibiting the learning and memory potassium channel activity as the active ingredient which can be obtained by the aforementioned screening method of the present invention.

Abstract: A method of detecting rpoB sequences of Mycobacterium tuberculosis present in a biological sample that includes steps of amplifying the M. tuberculosis rpoB sequence in vitro in a nucleic acid amplification mixture that includes specific disclosed primer sequences, and detecting the amplified sequences using multiple probes that provide sequence information by their specific hybridization to portions of the amplified nucleic acid. Compositions for amplifying and detecting in vitro the rpoB sequences of M. tuberculosis in a sample are disclosed.

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: A method for producing a monkey-derived embryonic stem cell comprising the steps of carrying out fertilization by insemination by in vitro fertilization or intracytoplasmic sperm injection using a monkey ovum and monkey sperms, thereby giving a fertilized ovum, allowing the fertilized ovum to differentiate into a blastocyst by in vitro culture, and establishing an ES cell line using the blastocyst; the monkey ES cell obtained by the method, a method for screening a reagent for specific differentiation into cell or tissue by using the ES cell; and a differentiated cell or differentiated tissue each differentiated from the ES cell. According to the present invention, applications of the embryonic stem cells to embryological studies clinical applications, experimental models, and the like on primates, studies of diseases, are expected.

Abstract: The methods and compositions of the present invention find use in altering cardiac-preferred expression in transgenic animals. The compositions of the invention include isolated nucleic acid molecules, expression cassettes, animal cells, transgenic animals, and transgenic mice. The transgenic animals of the invention exhibit inducible cardiac preferred expression of a nucleotide sequence of interest. The methods allow generation of transgenic animals with altered cardiac preferred expression of the nucleotide sequence of interest. In particular, the invention provides a method for altering the susceptibility of a transgenic animal to cardiopathy. A transgenic animal of the invention finds use in identifying anti-cardiopathic compounds.

Abstract: Viral vectors and methods of making such vectors are described that preferentially kill neoplastic but not normal cells, the preferred vector being an adenovirus that has the endogenous promoters in the E1A and/or E4 regions substituted with a tumor specific promoter which is preferably E2F responsive.

Abstract: Disclosed is the isolation and characterization of EI24, a gene whose 2.4 kb mRNA is induced following etoposide treatment. Induction of EI24 mRNA by etoposide required expression of wild-type p53. Overexpression of functional p53 was sufficient to induce expression of the EI24 mRNA. The EI24 mRNA was also induced in a p53-dependent manner by ionizing irradiation of primary murine thymocytes. The invention is thus directed to an isolated EI24 protein, nucleotide sequences coding for and regulating expression of the protein, antibodies directed against the protein, and recombinant vectors and host cells containing the genetic sequences coding for and regulating the expression of the protein sequence. Antibodies can be used to detect EI24 in biological specimens, including, for example, human tissue samples.

Abstract: The invention relates to a method for the stable inversion of a DNA fragment upon recombinase-mediated rearrangements using two sets of two incompatible site-specific recombinase targeting sites (SSRTS) in the same order but in reverse orientation flanking the DNA fragment to be inverted. The invention also relates to a method for the stable inversion of the DNA fragment upon rearrangement mediated by a recombinase such as Cre recombinase. The invention also relates to a method for obtaining a transgenic cell of which at least one allele of a DNA sequence of interest is invalidated by a process of conditional deletion and the genome of which has a reporter gene inserted at the place of the DNA fragment deleted by the process of conditional deletion. A method to generate targeting sites to perform site-specific recombination mediated cassette exchange is also provided.

Abstract: A method of identifying a genotype of a transgenic mouse and reducing the variability of a transagene expression in the transgenic mouse comprises the step of introducing into a mouse (a) a transgene expression cassette, (b) a coat color expression cassette and (c) an insulator positioned at 5? or 3? end of the transgene expression cassette.

Abstract: An antibiotic-independent vector for stable vector maintenance and high protein expression and a method for gene expression using this vector are disclosed. The stable maintenance of the vector is due to expression of a glutamic acid racemase to complement a D-glutamic acid auxotroph. No antibiotics, such as ampicillin, are necessary for the stable maintenance of this vector.

Abstract: Methods for introducing at least one gene encoding a product into at least one target cell of a mammalian host for use in treating the mammalian host are disclosed. These methods include employing recombinant techniques to produce a vector molecule that contains the gene encoding for the product, and infecting the target cells of the mammalian host using the DNA vector molecule. A method to produce an animal model for the study of connective tissue pathology is also disclosed.

Abstract: The present invention is directed generally to activating gene expression or causing over-expression of a gene by recombination methods in situ. The invention also is directed generally to methods for expressing an endogenous gene in a cell at levels higher than those normally found in the cell. In one embodiment of the invention, expression of an endogenous gene is activated or increased following integration into the cell, by non-homologous or illegitimate recombination, of a regulatory sequence that activates expression of the gene. The invention also provides methods for the identification, activation, isolation, and/or expression of genes undiscoverable by current methods since no target sequence is necessary for integration. Thus, by the present invention, endogenous genes, including those associated with human disease and development, may be activated and isolated without prior knowledge of the sequence, structure, function, or expression profile of the genes.

Abstract: A method for producing recombinant adeno-associated virus in the absence of contaminating helper virus or wild-type virus involves culturing a mammalian host cell containing an rAd/AAV hybrid virus, an AAV rep sequence and an AAV cap sequence under the control of regulatory sequences directing expression thereof. The rAd/AAV hybrid virus contains a rAAV construct to be packaged into an AAV virion in an backbone containing the adenoviral sequences necessary to express E1a and E1b gene products and to permit replication of the hybrid virus. The method of the invention permits replication of the hybrid virus and production of rAAV virion in this host cell in the absence of a helper virus and obviates a subsequent purification step to purify rAAV from contaminating virus.

Abstract: Viral vectors and methods of making such vectors are described that preferentially kill neoplastic but not normal cells, the preferred vector being an adenovirus that has the endogenous promoters in the E1A and/or E4 regions substituted with a tumor specific promoter which is preferably E2F responsive.

Abstract: The present invention discloses that DARPP-32 is substrate for the cyclin dependent kinase Cdk5. The phosphorylation takes place at a specific threonine residue of DARPP-32 (Threonine 75). The Cdk5 catalyzed phosphorylation of DARPP-32 converts this protein into an inhibitor of the cAMP dependent protein kinase (PKA) and furthermore prevents it from being converted to an inhibitor of protein phosphatase 1 (PP1). Methods of identifying agents that modulate the phosphorylation of DARPP-32 by Cdk5 are disclosed. Methods of treating dopamine dysfunction in animal subjects are also provided.

Abstract: The present invention relates to mammalian staufen, a double-stranded RNA-binding protein involved in mRNA transport and localization. The invention further relates to the demonstration of the association of a RNA-binding protein with the rough endoplasmic reticulum (RER), implicating staufen and related proteins in the transport of RNA to its site of translation. Broadly, the invention therefore relates to transport and translation of RNA. More specifically, the present invention relates to human and mouse staufen proteins and to the modulation of transport of RNA to the RER by these proteins. The present invention also relates to isolated nucleic acid molecules encoding mammalian staufen, as well as vectors and host cells harboring same. In addition, the present invention relates to screening assays for identifying modulators of staufen activity and to the identification of mutants thereof which abrogate their interaction with RER.

Abstract: A method for the preparation of an antisense oligonucleotide or derivative thereof comprising the steps of: selecting a target nucleic acid, if necessary elucidating its sequence; generating the antisense oligonucleotide with the proviso that: the oligonucleotide comprises at least 8 residues; the oligonucleotide comprises at maximum twelve elements, which are capable of forming three hydrogen bonds each to cytosine bases; the oligonucleotide does not contain four or more consecutive elements, capable of forming three hydrogen bonds each with four consecutive cytosine bases (CCCC) within the target molecule or alternatively four or more consecutive elements of GGGG; the oligonucleotide does also not contain 2 or more series of three consecutive elements, capable of forming three hydrogen bonds each with three consecutive cytosine bases (CCC) within the target molecule, or alternatively 2 or more series of three consecutive elements of GGG; and the ratio between residues forming two hydrogen bonds per residue

Abstract: More efficient transfer of genes into host cells or embryos to transform the cells or embryos is facilitated by transposition vectors using the minimal amount of nucleotide sequences in the transposon piggyBac required for gene transfer. The transformed cells or embryos may be developed into transgenic organisms.

Abstract: The present application discloses the preparation and use of attenuated tumor-targeted bacteria vectors for the delivery of one or more primary effector molecule(s) to the site of a solid tumor. The primary effector molecule(s) of the invention is used in the methods of the invention to treat a solid tumor cancer such as a carcinoma, melanoma, lymphoma, or sarcoma. The invention relates to the surprising discovery that effector molecules, which may be toxic when administered systemically to a host, can be delivered locally to tumors by attenuated tumor-targeted bacteria with reduced toxicity to the host. The application also discloses to the delivery of one or more optional effector molecule(s) (termed secondary effector molecules) which may be delivered by the attenuated tumor-targeted bacteria in conjunction with the primary effector molecule(s).