Abstract: The invention relates to methods of introducing a heterologous DNA sequence into a mouse embryonic stem cell wherein the DNA sequence is inserted by homologous recombination into a villin gene/I-SceI hybrid by creating a double strand break with I-SceI meganuclease. Subsequently, the mouse embryonic stem cells can be used to generate a transgenic mouse comprising the heterologous DNA sequence. Additionally, the methods can be used for gene replacement in ovo where a mouse oocyte containing a villin gene/I-SceI hybrid within its genome exists or is first generated. More generally, the methods can be used for the targeted insertion of a heterologous DNA sequence into any cell containing a villin gene/I-SceI hybrid sequence within its genome.

Abstract: The invention relates to a transgenic non-human mammal whose cells express a constitutively active oncogenic mutant of the kinase-domain of the Raf-1 gene or a protein coded by a corresponding normal allele or derivative of the A, B or c-Raf-1 gene.

Abstract: A gene is transferred to the early stage chicken embryo by electroporation with placing the electrode tips to hold both ends of the embryo. Transfer of the desired gene to the early stage embryo was detected using the &bgr;-galactosidase gene of E coli as a reporter gene. The expression of the gene in the early stage embryo of chicken was thus confirmed. This method for transferring a gene into undifferentiated cells is simple to implement, highly efficient in transferring a gene, and applicable to various animal species.

Abstract: The invention provides a transgenic non-human animal expressing a perlecan encoding transgene. Also provided is a double-transgenic non-human animal expressing a perlecan and a amyloid encoding transgene. A method of screening for a compound which alters the rate or extent of amyloid deposition is additionally provided. The method consists of: (a) constructing a perlecan transgenic animal; (b) administering an effective amount of a test compound to said perlecan transgenic animal; and (c) determining whether said test compound alters the extent or rate of amyloid deposition. Finally, the invention provides a method of screening for a compound which alters the rate or extent of amyloid deposition. The method consists of: (a) constructing a perlecan/amyloid double-transgenic animal; (b) administering an effective amount of a test compound to said perlecan/amyloid double-transgenic animal; and (c) determining whether said test compound alters the extent or rate of amyloid deposition.

Abstract: The present invention provides a gene search method comprising P-element insertion and being capable of efficiently identifying a novel gene regulating various biological functions of Drosophila and specifying a function corresponding to the novel gene. An unknown gene of Drosophila can be searched by the method comprising a step of inserting in the genome of Drosophila a gene search vector carrying two sets of an expression regulatory sequence comprising UAS sequence for GAL4 transcription factor GAL4 and a promoter sequence, as integrated in the P-element sequence in such a manner that their downstreams are in opposite directions, mating the vector-inserted Drosophila with a Drosophila expressing the GAL4 to create progeny individuals, and identifying a vector-inserted line with a phenotype different from those of wild-type Drosophila, and determining the nucleotide sequence of the gene for the mutant phenotype.

Abstract: A mammal lacking expression of particular CD45 isoform in certain cells of the immune system is provided. The mammal may optionally contain a transgene encoding the CD45RO isoform. Also provided are methods of using these mammals.

Abstract: Vectors and methods are provided for introducing genetic material into cells of a chicken or other avian species. More particularly, vectors and methods are provided for transferring a transgene to an embryonic chicken cell, so as to create a transgenic hen wherein the transgene is expressed in the hen's oviduct and the transgene product is secreted in the hen's eggs and/or those of her offspring. In a preferred embodiment, the transgene product is secreted in the egg white.

Abstract: The present invention provides improved methods and compositions for the generation of transgenic non-human animals. The present invention permits the introduction of exogenous nucleic acid sequences into the genome of unfertilized eggs (e.g., pre-maturation oocytes and pre-fertilization oocytes) by microinjection of infectious retrovirus into the perivitelline space of the egg. The methods of the present invention provide an increased efficiency of production of transgenic animals with a reduced rate of generating animals which are mosaic for the presence of the transgene.

Abstract: The invention relates to a method of identifying nuclear receptor controlled genes in specific tissues. In particular, the method also provides a method of activating PPAR&ggr; nuclear receptor controlled target genes in vivo in a tissue-specific manner.

Abstract: This invention relates to a mouse with deficiency of gene of neutrophil chemotactic factor LECT2 having a function of lacking by deleting a whole or a part LECT2 gene or inserting other gene such as a selective marker gene to any portion of the LECT2 gene or replacing it with another gene(s).

Abstract: The invention provides novel polypeptides which are associated with the transcription complex NF-AT, polynucleotides encoding such polypeptides, antibodies which are reactive with such polypeptides, polynucleotide hybridization probes and PCR amplification probes for detecting polynucleotides which encode such polypeptides, transgenes which encode such polypeptides, homologous targeting constructs that encode such polypeptides and/or homologously integrate in or near endogenous genes encoding such polypeptides, nonhuman transgenic animals which comprise functionally disrupted endogenous genes that normally encode such polypeptides, and transgenic nonhuman animals which comprise transgenes encoding such polypeptides.

Abstract: The present invention pertains to a method for treating obesity in a mammal which comprises reducing the biological activity of HMGI genes in the mammal. In another embodiment, the invention pertains to a method for treating a tumor in a patient by reducing the biological activity of normal HMGI genes which comprises administering to the patient a therapeutically effective amount of an inhibitor compound active against normal HMGI-C or HMGI(Y) genes. In another embodiment, the invention pertains to a method of producing a transgenic non-human mammal, the germ cells and somatic cells of which contain an inactivated HMGI gene sequence introduced into the mammal at an embryonic stage. In another embodiment, the invention pertains to a method for screening candidate compounds capable of inhibiting the biological activity of normal HMGI proteins. In another embodiment, the invention pertains to a method for screening candidate compounds capable of inhibiting the biological activity of normal HMGI genes.

Abstract: The present invention relates to human bile salt-stimulated lipase (BSSL) obtainable from transgenic sheep. The invention further relates to transgenic sheep whose germ cells and somatic cells contain a recombinant nucleotide molecule comprising a nucleotide sequence encoding for human BSSL. The invention also relates to methods for producing said transgenic animals, as well as to methods for producing human BSSL derived from transgenic animals. In addition, the invention provides the use of compositions comprising BSSL in the treatment of diseases relating to exocrine pancreatic insufficiency, and for improvement of the utilization of dietary lipids in preterm born infants.

Abstract: The present invention relates generally to the fields of oligonucleotide synthesis. More particularly, it concerns the assembly of genes and genomes of completely synthetic artificial organisms. Thus, the present invention outlines a novel approach to utilizing the results of genomic sequence information by computer directed gene synthesis based on computing on the human genome database. Specifically, the present invention contemplates and describes the chemical synthesis and resynthesis of genes defined by the genome sequence in a host vector and transfer and expression of these sequences into suitable hosts.

Abstract: Non-human mammalian animals having a higher epidermal expression level of protein kinase C&egr; than their wild-type counterparts are phenotypically distinguished from wild-type animals in that the animals induced to develop tumors in a chemical initiation/promotion protocol are suppressed for subsequent papilloma development but are susceptible to developing squamous cell carcinoma and metastatic squamous cell carcinoma. The animals are advantageously used in methods for screening putative agents for altering the susceptibility, development and progression of squamous cell carcinoma and metastatic squamous cell carcinoma and have further commercial value as tools for investigating the development of metastatic disease.

Abstract: A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand's Factor into milk may be used to stabilize newly-secreted Factor VIII.

Abstract: A transgenic rabbit which has in its genomic DNA sequences that encode apolipoprotein (a) and apolipoprotein B polypeptides which are capable of combining to produce lipoprotein (a), a process for creating such a rabbit, and the use of the rabbit to identify compounds which are effective in the treatment of human diseases which are associated with, induced and/or exacerbated by Lp(a) expression.

Abstract: The present invention features a non-human animal model that is susceptible to infection by human hepatotrophic pathogens, particularly human hepatitis C virus (HCV). The model is based on a non-human, immunocompromised xenogeneic transgenic animal having a human-mouse chimeric liver, where the transgene provides for expression of a urokinase-type plasminogen activator in the liver. The invention also features methods for identifying candidate therapeutic agents, e.g., agents having antiviral activity against HCV infection.

Abstract: Animal model useful for testing potential therapeutic agents for the treatment of neurodegenerative disorders, in particular disorders associated with the presence of Lewy pathology.

Abstract: The present invention provides methods which greatly facilitate the rapidity in which cells and transgenic animals with targeted genes may be generated. The invention hastens the investigation of cells and transgenic animals bearing lowered expression of the targeted gene product, a truncated targeted gene product, a fusion protein of the targeted gene and exogenous DNA, or the expression of a different gene from the locus of the targeted gene whose product has reduced expression levels. Also disclosed is a transgenic animal having Cushing's disease. Also disclosed are diagnostic methods for detecting patients with endocrine disorders, and methods for treating or alleviating the symptoms of endocrine disorders.

Abstract: A transgenic large mammal is produced by a method including the steps of obtaining one or more early embryos, selectively preparing an embryo having at least three cells, and preferably at a stage in development corresponding in time to the onset of transcription of the embryo's paternal genome, and introducing isolated nucleic acid molecules into a blastomere of the selected embryo. The introduction of isolated nucleic acid molecules into such embryos results in the generation of transgenic large mammals at a significantly increased frequency as compared to introducing isolated nucleic acid molecules into zygotes or into the blastomeres of embryos at the one or two cell stage of development.

Abstract: Transgenic mice are constructed by binding the “hHB-EGF/DTR” gene to the downstream of an albumin enhancer/promoter that is expressed specifically in hepatic parenchymal cells and introducing this unit into mice. After the “hHB-EGF/DTR” gene has been confirmed to be expressed specifically in hapatic cells, diphtehria toxin is administered to the transgenic mice to examine whether the hepatic parenchymal cells are disrupted. The hepatic cells of the transgenic mice can be selectively dirupted depending on the administration period of the diphtheria toxin.

Abstract: One of the problems facing transplantation procedures today is the availability of organs suitable for transplantation. A way around the lack of organs, it to harvest organs from animals. However, immune reactions between discordant donor and recipient mammals presents a major obstacle in such transplantation procedures. The production of transgenic non-human mammals expressing a DNA sequence encoding an homologous complement restriction factor or factors of a discordant mammalian species such that the expression of the DNA sequence would prevent xenograft rejection by the host mammal's immune system would provide organs, tissues and cells for transplantation.

Abstract: Process for producing mammals with defined genetic properties, particularly transgenic mammals. Animals which are derived completely from ES cells are obtained by injecting totipotent cells (embryonic stem cells or embryonic germ cells) genetically manipulated and cultured in vitro into tetraploid blastocysts and implanting the resulting embryo in a foster mother, in a single operation.

Abstract: Process for producing mammals with defined genetic properties, particularly transgenic mammals. Animals which are derived completely from ES cells are obtained by injecting totipotent cells (embryonic stem cells or embryonic germ cells) genetically manipulated and cultured in vitro into tetraploid blastocysts and implanting the resulting embryo in a foster mother, in a single operation.

Abstract: The present invention provides mice which are deficient in the normal expression of one or more members of the RAR or RXR class of receptors, to mice heterozygous for such deficiency, and to cell lines, preferably pluripotent or totipotent cell lines, which are heterozygous or homozygous for such deficiency. The present invention further provides the use of any of the above mice and cell lines in situations where the absence of at least one RAR or RXR receptors, or the normal expression thereof, is desirable.

Abstract: Transgenic nonhuman mammals, such as transgenic mice, which lack erythropoietin expression, in which the erythropoietin receptor is deleted, which carry a heterologous erythropoietin receptor (e.g., a chimeric receptor); constructs useful for producing such transgenic nonhuman mammals, embryonic stem cells containing the constructs, a method of producing the transgenic nonhuman mammals and a method of identifying erythropoietin mimics or mimetics.

Abstract: The present invention involves the role of p53 in the differentiation of embryonic tissues. More particularly, the present invention provides methods of the blocking of p53 function in embryonic tissues, and the use of these tissues as screening tools for substances that are capable of overcoming the p53-related block in differentation, both in vitro and in vivo. The similarities between undifferentiated embryonic cells and tumor cells is evident, and thus these assays serve as a model for possible cancer therapeutics. In addition, methods for identifying additional cellular components that interact p53 or p53-related pathways are provided.

Abstract: The present invention provides transgenic fish whose somatic and germ cells contain a genomically integrated plasmid containing a heterologous mutation target nucleic acid sequence that is detectable via bioassay in a bacterial cell into which the target nucleic acid has been introduced. The frequency and character of mutations in the mutatable target nucleic acid sequence following exposure of the transgenic fish to one or more potentially mutagenic agents can thus be evaluated.

Abstract: The present invention provides for genetically engineered non-human animals, including but not limited to mice, which lack one or more endogenous neurofilament gene. Human neurofilament genes may be introduced into such genetically non-human animals (referred to hereafter as “knockout animals”) to produce improved models of the physiology of human neurofilament proteins which may be used to study human neurofilament-associated neurodegenerative conditions.

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 being characterized by the ability to hybridize to the DNA sequence of Minos 1 under stringent hybridization conditions. The invention also relates to a purified transposase protein, or peptide fragments thereof, encoded by such DNA sequences. Such transposable are useful in methods for 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 Tc-1 family of transposable elements.

Abstract: The present invention relates to a method for transfection of avian primordial germ cells which comprises a step of electroporating a foreign gene into avian primordial germ cells after addition of dimethylsulfoxide( “DMSO”) to culture medium containing the cells, and a method for production of transgenic Aves by employing the avian primordial germ cells transfected thereby. The DMSO-electroporation method of the invention has various advantages over the conventional transfection methods in light of high transfection efficiency, safety, and simple manipulation. Accordingly, a large number of primordial germ cells transfected with a gene of interest can be efficiently produced by the invention, which, in turn, may increase the rate of transfer of exogenous DNA into recipient embryos.

Abstract: The present invention relates to a method of identifying drugs or agents which have immuno-suppressive effects through or as a result of their effect on calcineurin, including drugs which affect the calcineurin A&agr; (CNA&agr;) subunit or the calcineurin A&bgr; (CNA&bgr;) subunit. In addition, the present invention relates to a method of identifying drugs which reduce (partially or totally) phosphorylation of the microtubule-associated protein tau, in the nervous system of a mammal; a method of identifying drugs which reduce (partially or totally) paired helical filament formation in the nervous system of a mammal; and a method of identifying drugs which reduce (partially or totally) formation of paired helical filaments, amyloid deposits or both. The present invention also relates to transgenic non-human mammals, such as rodents and particularly mice, which lack a functional calcineurin gene and, thus, have disyrupted calcineurin expression.

Abstract: An animal, e.g., transgenic mouse, in which the MSH5 gene is misexpressed. The animal is useful for screening treatments for a number of conditions. Methods for identifying contraceptive agents are also described.

Abstract: The invention provides methods and compositions for defining a cell type, generally involving the steps of (a) amplifying the mRNA of a single cell of a heterogenous population of cells; (b) probing a comprehensive expression library with the amplified mRNA to define a gross expression profile of the cell; and (c) comparing the gross expression profile of the cell with a gross expression profile of one or more other cells to define a unique expression profile of the cell, wherein the unique expression profile of the cell provides a marker defining the cell type

Abstract: A hepatitis type C animal model into which cDNA derived from hepatitis C virus has been introduced. This animal model is useful for clarification of an onset mechanism of hepatitis C and as well as for development of means for treating the disease.

Abstract: A transgenic non-human animal with alterations in the osteopontin gene is prepared by introduction of a gene encoding an altered osteopontin protein into a host non-human animal. Methods for using transgenic mice so generated to screen for agents that effect osteopontin's cellular modulating activity are also provided.

Abstract: A transgenic mammal whose somatic and germ cells having a nucleic acid construct wherein the construct includes a mammalian promoter operably linked to a cDNA genomic sequence is provided for the overexpression of galanin. Also provided is a construct having cDNA for the overexpression of galanin. A method of making a transgenic mammal by producing a mammal having a construct for the overexpression of galanin is provided.

Abstract: Switch regions derived from an immunoglobulin (Ig) gene are used to direct recombination between a targeting construct containing a promoter, a switch region (S1), and 2) a target locus minimally containing a promoter, a switch region (S2), and a target sequence.

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: This invention relates to methods and compositions of controlling cell distribution within a bioartificial organ by exposing the cells to a treatment that inhibits cell proliferation, promotes cell differentiation, or affects cell attachment to a growth surface within the bioartificial organ. Such treatments include (1) genetically manipulating cells, (2) exposing the cells to a proliferation-inhibiting compound or a differentiation-inducing compound or removing the cells from exposure to a proliferation-stimulating compound or a differentiation-inhibiting compound; exposing the cells to irradiation, and (3) modifying a growth surface of the BAO with ECM molecules, molecules affecting cell proliferation or adhesion, or an inert scaffold, or a combination thereof. These treatments may be used in combination.

Abstract: Transgenic mice having a phenotype characterized by the substantial depletion of a mature lymphocytic cell type otherwise naturally occurring in the species from which the transgenic mouse is derived. The phenotype is conferred in the transgenic mouse by a transgene contained in at least the precursor stem cell of the lymphocytic cell type which is depleted. The transgene comprises a DNA sequence encoding a lymphatic polypeptide variant which inhibits maturation of the lymphocytic cell type.

Abstract: Disclosed are transgenic fish, and a method of making transgenic fish, which express transgenes in stable and predictable tissue- or developmentally-specific patterns. The transgenic fish contain transgene constructs with homologous expression sequences. Also disclosed are methods of using such transgenic fish. Such expression of transgenes allow the study of developmental processes, the relationship of cell lineages, the assessment of the effect of specific genes and compounds on the development or maintenance of specific tissues or cell lineages, and the maintenance of lines of fish bearing mutant genes.

Abstract: A gene is transferred to the early stage chicken embryo by electroporation with placing the electrode tips to hold both ends of the embryo. Transfer of the desired gene to the early stage embryo was detected using the &bgr;-galactosidase gene of E coli as a reporter gene. The expression of the gene in the early stage embryo of chicken was thus confirmed. This method for transferring a gene into undifferentiated cells is simple to implement, highly efficient in transferring a gene, and applicable to various animal species.

Abstract: Disclosed herein are an expression vector of mud loach growth hormone gene and a fast-growing transgenic mud loach transformed with the expression vector. Also disclosed herein is an expression vector containing &bgr;-actin gene regulation site of mud loach which is constructed for expression of useful genes in fishes. The transgenic mud loaches transformed with the expression vector of mud loach growth hormone gene show growth rate of 25 times higher than that of normal mud loaches.

Abstract: The present invention provides a transgenic non-human mammal which lacks a functional UCP2 gene. The UCP-2 deficient transgenic knockout mammal described herein provides a source of cells and animals useful to practice methods for the identification and/or evaluation of agents for their ability to affect signaling in cells, such as pancreatic &bgr;-cells, in which ATP serves a regulatory function. Further aspect of the invention provide a method for the identification of agents (e.g., therapeutic agents) which inhibit UCP2 activity; a method for the identification of agents which mimic UCP2 activity and a method of treating diseases or conditions associated with UCP2 function (e.g., negative regulation or uncoupling activity).

Abstract: The present invention is based on the generation and phenotypic characterization of transgenic knockout homozygous rchd534 mutant mice which display characteristic cardiovascular disease symptoms. Such transgenic knockout homozygous rchd534 mutant mice are useful models for the analysis and characterization of rchd534 protein involvement in development and homeostasis of the cardiovascular system and tissue-specific regulation of the TGF-&bgr; signaling pathways. Such transgenic mice may be used for screening compounds that may potentially useful for treating or preventing cardiovascular disease.

Abstract: The present invention provides a gene search method comprising P-element insertion and being capable of efficiently identifying a novel gene regulating various biological functions of Drosophila and specifying a function corresponding to the novel gene. An unknown gene of Drosophila can be searched by the method comprising a step of inserting in the genome of Drosophila a gene search vector carrying two sets of an expression regulatory sequence comprising UAS sequence for GAL4 transcription factor GAL4 and a promoter sequence, as integrated in the P-element sequence in such a manner that their downstreams are in opposite directions, mating the vector-inserted Drosophila with a Drosophila expressing the GAL4 to create progeny individuals, and identifying a vector-inserted line with a phenotype different from those of wild-type Drosophila, and determining the nucleotide sequence of the gene for the mutant phenotype.

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 being characterized by the ability to hybridize to the DNA sequence of Minos 1 under stringent hybridization conditions. The invention also relates to a purified transposase protein, or peptide fragments thereof, encoded by such DNA sequences. Such transposable are useful in methods for 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 Tc-1 family of transposable elements.

Abstract: The present invention relates to a transgenic non-human animal embryo lacking native presenilin 1 and a transgenic non-human animal having only a non-native presenilin 1. The transgenic animals and cells derived therefrom can be used in the study of the expression pattern, activity and modulators of presenilin 1, in the study of the role of presenilin 1 in Alzheimer's Disease and in the study of disorders of the central nervous system.