Abstract: A method of producing a cloned or genetically modified non-human mammalian embryo comprising: (a) providing a cell culture comprising a plurality of in vitro matured oocytes; (b) preferentially selecting from the cell culture a rapidly matured oocyte or developmentally competent oocyte; (c) transferring DNA from a donor cell derived from non-human mammalian tissue to the matured oocyte to form a nuclear transfer unit; and (d) culturing said nuclear transfer unit to form an embryo. At the initiation of maturation, oocytes are preferably beyond the GV-II stage of prophase I. Porcine oocytes most preferably mature in about 20-28 hours.

Abstract: The invention discloses chimeric milk-producing tissues containing human mammary cells implanted into cleared mammary fat pad tissue or other suitable tissue of a non-human animal host, and discloses the use of human milk produced by chimeric milk-producing tissues. The invention further provides methods for avoiding problems of xenogeneic transplantation in chimeric milk-producing tissues.

Abstract: A nuclear transfer method is provided wherein nuclear DNA in whole or part is injected into enucleated oocytes. The method is suitable for different donor cells, and preferably ES cells.

Abstract: The present invention provides data to demonstrate that the fusion performance of a cell-line in procedures involving fusion and cleavage indices either alone or in combination are a means for selecting a cell lines that will be successful in a nuclear transfer or microinjection program. This technique and method of selecting a cell line offers an additional alternative and improvement in the creation of activated and fused nuclear transfer-capable embryos for the production of live offspring in various mammalian non-human species including goats, pigs, rodents, primates, rabbits and cattle.

Abstract: Transgenes for producing recombinant polypeptides transgenic bovine species. A transgene for producing recombinant polypeptides in the milk of transgenic bovine species comprises at least one expression regulation sequence, a secretory DNA sequence encoding a secretory signal sequence which is functional in mammary secretory cells of the bovine species and a recombinant DNA sequence encoding the recombinant polypeptide. Also included are methods for producing transgenic bovine species. The method includes introducing the above transgene into an embryonal target cell of a bovine species, transplanting the transgenic embryonic target cell formed thereby into a recipient bovine parent and identifying at least one female offspring which is capable of producing the recombinant polypeptide in its milk.

Abstract: Selection and cloning methods are disclosed that are effective for the selective multiplication of desired animals, for instance livestock animals. These methods can be used to expand populations of animals, and are particularly useful for duplicating animals selected based on traits that are measured after the animal is deceased. Certain embodiments include techniques useful for selecting a desirable bovine animal (e.g., desirable steer) based at least in part on a measurable carcass trait. Also described are specific cloning techniques that involve repetitive (for instance, two) cycles of cloning, such as nuclear transfer cloning. In specific embodiments, a two-step cloning system is described, in which the nuclear donor of the first cloning cycle is an adult fibroblast cell and the nuclear donor of the second cloning cycle is a fetal fibroblast harvested from a fetus that arose from the list cloning cycle.

Abstract: A process for the production of a peptide is disclosed, the process comprising expressing in the milk of a transgenic, non-human, placental mammal a fusion protein which comprises the peptide to be expressed linked to a fusion partner protein which is lysozyme. The fusion protein may be separate from the milk and cleaved to yield the target peptide. A transgenic, non-human, placental mammal whose genome incorporates a DNA molecule comprising a coding sequence encoding lysozyme coupled to a peptide is also described.

Abstract: A transgenic non-human animal of the species selected from the group consisting of avian, bovine, ovine and porcine having a transgene which results in disrupting the production of and/or activity of growth differentiation factor-11 (GDF-11) chromosomally integrated into the germ cells of the animal is disclosed. Also disclosed are methods for making such animals, and methods of treating animals, including humans, with antibodies or antisense directed to GDF-11. The animals so treated are characterized by increased muscle tissue and bone tissue.

Abstract: Nucleic acid and protein sequences relating to a gene required for systemic RNAi are disclosed. The SID-1 protein is shown to be required for systemic RNAi. Nucleic acids, vectors, transformed cells, transgenic animals, polypeptides, and antibodies relating to the sid-1 gene and protein are disclosed. Also provided are methods for reducing the expression of a target gene in a cell, a population of cells, or an animal.

Abstract: The invention generally relates to synthesis of essential fatty acids and their derivatives, long-chain polyunsaturated fatty acids (LC-PUFAs) and eicosanoids in transfected cells and in transgenic animals.

Abstract: The present invention provides animal model systems for cartilage-degenerative disease, which comprise transgenic animals which can express recombinant matrix-degrading enzymes (MDEs), particularly matrix metalloproteinases (MMPs), in a temporally and spatially regulated manner. The invention also provides methods for producing phenotypic indicators of cartilage-degenerative disease in a mammal and methods for determining the potential of a composition to counteract cartilage-degenerative disease. The invention also provides isolated nucleic acids encoding proMMP polypeptides that exhibit constitutive enzymatic activity and isolated proMMP polypeptides.

Abstract: Nuclear transfer methods and techniques involving cryopreserved bovine somatic cells, nuclei, or nuclear DNA, and the products thereof, are disclosed. These methods employ an enucleated oocyte as a recipient, which is fused with a donor nuclear genome from a cryopreserved bovine somatic cell to form a couplet. In addition, the a cloned bull exhibiting disease resistance is disclosed.

Abstract: The present invention relates to cloning technologies. The invention relates in part to immortalized and totipotent cells useful for cloning animals, the embryos produced from these cells using nuclear transfer techniques, animals that arise from these cells and embryos, and materials, methods, and processes for establishing such cells, embryos, and animals.

Abstract: This invention relates to a process for breeding animals through cloning as well as animals obtainable with the process, in particular to a process for the reproduction of animal embryos via an efficient nucleus transfer with foetal fibroblasts.

Abstract: Disclosed is a non-human mammal comprising a cell that comprises an expression vector containing a promoter and a PTTG carboxy-terminal-related DNA.

Abstract: The invention provides transgenic nonhuman mammals expressing C1 inhibitor in their milk. The C1 inhibitor is useful in treating patients with hereditary angioedema or patients requiring immunosuppression.

Abstract: The present invention provides for a transgenic nonhuman mammal whose germ or somatic cells contain a nucleic acid molecule which encodes calcineurin or a variant thereof under the control of a regulatable promoter, introduced into the mammal, or an ancestor thereof, at an embryonic stage. The present invention also provides for a method of evaluating whether a compound is effective in improving long-term memory in a subject suffering from impaired long-term memory which comprises: (a) administering the compound to the transgenic nonhuman mammal of claim 1 wherein the mammal has increased brain-specific calcineurin activity due to expression of the nucleic acid, and (b) comparing the long-term memory of the mammal in step (a) with the long-term memory of the mammal in the absence of the compound so as to determine whether the compound is effective in rescuing the long-term memory defect in the subject.

Abstract: Disclosed is a non-human mammal comprising a cell that comprises an expression vector containing a promoter and a PTTG carboxy-terminal-related DNA.

Abstract: A gene (cDNA) encoding a bovine myostatin protein. The nucleic acid coding sequence is identified as SEQ ID NO:1 and the protein sequence is identified as SEQ ID NO:2. A mutant gene (SEQ ID NO:3) in which the coding sequence lacks an 11-base pair consecutive sequence (SEQ ID NO:11) of the sequence encoding bovine protein having myostatin activity has been sequenced. It has been shown that cattle of the Belgian Blue breed homozygous for the mutant gene lacking myostatin activity are double-muscled. A method for determining the presence of muscular hyperplasia in a mammal is described. The method includes obtaining a sample of material containing DNA from the mammal and ascertaining whether a sequence of the DNA encoding (a) a protein having biological activity of myostatin, is present, and whether a sequence of the DNA encoding (b) an allelic protein lacking the activity of (a), is present. The absence of (a) and the presence of (b) indicates the presence of muscular hyperplasia in the mammal.

Abstract: The present invention provides a method for producing cloned cows by employing in vitro maturation of oocytes and nuclear transfer techniques. The method comprises collecting donor somatic cell lines from cows; maturing oocytes extracted from an ovary in vitro; enucleating the oocyte by cutting a portion of the zona pellucida and squeezing out a portion of the cytoplasm, including the first polar body; inserting the donor somatic cell into the oocyte; electrofusing the cells to produce embryos; post-activating the embryos; transferring them into surrogate cows to produce cloned calves.

Abstract: The present invention provides methods of producing transgenic livestock animals. The methods generally involve first introducing a nucleoprotein made up of nucleic acid and a recombinase into a totipotent or pluripotent cell to produce a recombinant totipotent or pluripotent cell and then growing the recombinant totipotent or pluripotent cell to produce the transgenic livestock animal. The invention further provides kits for use in generating transgenic non-human animals of the invention.

Abstract: This invention provides methods for reducing the amount of Dnmt3L in a cell, obtaining a cell having a reduced amount of Dnmt3L, and cells, tissues, organs and non-human transgenic mammals having a reduced amount of Dnmt3L. This invention further provides related methods for producing a mammalian zygote in vitro which, upon successful subsequent development in utero, gives rise to a mammal having a reduced susceptibility to an abnormality associated with epigenetic instability. This invention further provides methods for determining the amount of Dnmt3L in a cell, and methods for determining whether agents bind to Dnmt3L or decrease the amount of Dnmt3L in a cell. Finally, this invention provides related nucleic acids and compositions.

Abstract: The present invention concerns a method for in vivo generation of a linear polynucleotide with 5′ and 3′ free ends from a vector having no free end, said linear polynucleotide being integrated into the host cell genome. The vector having no free end according to the present invention comprise the polynucleotide to be linearized or excised flanked by a cleavage site, said cleavage site being preferably not found in the host cell genome. The present invention also relates to the resulting cells and their uses, for example for production of proteins or other genes, biomolecules, biomaterials, transgenic plants, vaccines, transgenic animals or for treatment or prophylaxis of a condition or disorder in an individual.

Abstract: A method of reconstituting an animal embryo involves transferring a diploid nucleus into an oocyte which is arrested in the metaphase of the second meiotic division. The oocyte is not activated at the time of transfer, so that the donor nucleus is kept exposed to the recipient cytoplasm for a period of time. The diploid nucleus can be donated by a cell in either the G0 or G1 phase of the cell cycle at the time of transfer. Subsequently, the reconstituted embryo is activated. Correct ploidy is maintained during activation, for example, by incubating the reconstituted embryo in the presence of a microtubule inhibitor such as nocodazole. The reconstituted embryo may then give rise to one or more live animal births. The invention is useful in the production of transgenic animals as well as non-transgenics of high genetic merit.

Abstract: A method of reconstituting an animal embryo involves transferring a diploid nucleus into an oocyte which is arrested in the metaphase of the second meiotic division. The oocyte is not activated at the time of transfer, so that the donor nucleus is kept exposed to the recipient cytoplasm for a period of time. The diploid nucleus can be donated by a cell in either the G0 or G1 phase of the cell cycle at the time of transfer. Subsequently, the reconstituted embryo is activated. Correct ploidy is maintained during activation, for example, by incubating the reconstituted embryo in the presence of a microtubule inhibitor such as nocodazole. The reconstituted embryo may then give rise to one or more live animal births. The invention is useful in the production of transgenic animals as well as non-transgenics of high genetic merit.

Abstract: The present invention relates to the stabilisation of milk from transgenic animals. In particular, the invention relates to the protection of proteins (e.g. fibrinogen) expressed in milk from transgenic animals by co-expression of a serine proteinase inhibitor (e.g. &agr;1-antitrypsin) in the milk of the transgenic animals.

Abstract: Described is a method for the production of a heteromeric protein comprising a first and a second peptide chain, wherein the first peptide chain is expressed in an animal, and the second peptide chain in another animal. The peptide chains are isolated from the animals and combined, resulting in formation of a functional protein. Optionally, both animals are mated, leading to offspring animals expressing both first and second peptide chains and producing the protein.

Abstract: Disclosed is a non-human mammal comprising a cell that comprises an expression vector containing a promoter and a PTTG carboxy-terminal-related DNA

Abstract: This invention provides bovine pregnancy test methods and devices. The test is also suitable for other ruminant and/or ungulate animals. Antigens from Group A (early pregnancy antigens), and/or Group B (mid-pregnancy antigens), and Group C (early, mid- and late pregnancy antigens) are detected in a fluid from the animal, and pregnancy is reliably determined. The pregnancy assays of this invention are preferably carried out using immunoassay devices which provide immediate results in the field.

Abstract: A transgenic non-human animal of the species selected from the group consisting of avian, bovine, ovine and porcine having a transgene which results in disrupting the production of and/or activity of growth differentiation factor-8 (GDF-8) chromosomally integrated into the germ cells of the animal is disclosed. Also disclosed are methods for making such animals, and methods of treating animals, including humans, with antibodies or antisense directed to GDF-8. The animals so treated are characterized by increased muscle tissue and bone content.

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 present invention relates to cloning technologies. The invention relates in part to immortalized and totipotent cells useful for cloning animals, the embryos produced from these cells using nuclear transfer techniques, animals that arise from these cells and embryos, and materials, methods, and processes for establishing such cells, embryos, and animals.

Abstract: The invention relates to an animal model of cancer. The animal carries a tumor xenograft and is immunosuppressed by administration of cyclosporin and ketoconazole. The model is useful for studying cancer and treatment thereof.

Abstract: The present invention relates to a method of producing an ungulate having both copies of the IgM heavy chain (mu) rag-1 and/or rag-2 gene eliminated from its genome. Animals which have IgM, rag-1 and/or rag-2 eliminated from their genome are unable to conduct the gene rearrangements that are necessary to generate the antigen receptors of B or T lymphocytes, and therefore will not develop native B or T cells. Because they are unable to produce B and T lymphocytes, these IgM, rag-1 or rag-2 ungulates cannot reject human hematopoietic stem cell preparations, and B and T lymphocytes which develop therefrom. Therefore, the present invention also involves injecting into IgM, rag-1 and/or rag-2 deficient ungulates, in utero or shortly after birth, human B and T lymphocytes whose immune systems produce human immunoglobulin that can be processed for therapeutic uses in humans.

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 cow cells in which a gene associated with mad cow disease has been modified to reduce susceptibility to mad cow disease, cows having reduced susceptibility to mad cow disease, nucleic acids for making such cells and cows, and products obtained from such cows. The invention also includes methods of making each of the foregoing.

Abstract: This invention relates to a method for breeding animals via cloning and the animals obtainable by the method, in particular a method for reproducing animal embryos via efficient nuclear transfer with primordial gametes.

Abstract: The invention relates to an animal model for studying behavior related to fatty acid amide and hydrolysis of fatty acid amide. The invention provides transgenic animals in which the protein fatty acid amide hydrolase is not expressed, and methods of using such animals.

Abstract: A method of reconstituting an animal embryo involves transferring a diploid nucleus into an oocyte which is arrested in the metaphase of the second meiotic division. The oocyte is not activated at the time of transfer, so that the donor nucleus is kept exposed to the recipient cytoplasm for a period of time. The diploid nucleus can be donated by a cell in either the G0 or G1 phase of the cell cycle at the time of transfer. Subsequently, the reconstituted embryo is activated. Correct ploidy is maintained during activation, for example, by incubating the reconstituted embryo in the presence of a microtubule inhibitor such as nocodazole. The reconstituted embryo may then give rise to one or more live animal births. The invention is useful in the production of transgenic animals as well as non-transgenics of high genetic merit.

Abstract: A method of reconstituting an animal embryo involves transferring a diploid nucleus into an oocyte which is arrested in the metaphase of the second meiotic division. The oocyte is not activated at the time of transfer, so that the donor nucleus is kept exposed to the recipient cytoplasm for a period of time. The diploid nucleus can be donated by a cell in either the G0 or G1 phase of the cell cycle at the time of transfer. Subsequently, the reconstituted embryo is activated. Correct ploidy is maintained during activation, for example, by incubating the reconstituted embryo in the presence of a microtubule inhibitor such as nocodazole. The reconstituted embryo may then give rise to one or more live animal births. The invention is useful in the production of transgenic animals as well as non-transgenics of high genetic merit.

Abstract: The present invention relates to the production of a transgenic bovine which comprises a genetic modification that results in inactivation and loss of expression of its endogenous antibodies, and the expression of xenogenous antibodies, preferably human antibodies. This is effected by inactivation of the IgM heavy chain expression and, optionally, by inactivation of the Ig light chain expression, and by the further introduction of an artificial chromosome which results in the expression of non-bovine antibodies, preferably human antibodies.

Abstract: An animal model is provided which is genetically engineered to express human serum albumin, and such animals may be advantageously used in assessing drugs, vaccines or other therapeutic compounds that may be used in humans. In addition, an animal model is provided which does not manufacture its own albumin and which has been injected with human serum albumin. Through the use of these animal models, drugs and other chemicals can be more accurately assessed in physiological environments that reflect the conditions to be expected in humans, and such models will be useful in assessing new drugs and evaluating toxic substances for potential dangers as carcinogens, mutagens, etc. Other applications include evaluating immunological properties of various albumin-engineered proteins which might be administered to humans as therapeutics or vaccines, and research of disease states, such as genetic diseases, to provide further insight in treating these diseases.

Abstract: Methods for preparing cell lines and transgenic animals that contain artificial chromosomes, methods for preparation of artificial chromosomes, methods for purification of artificial chromosomes, methods for targeted insertion of heterologous DNA into artificial chromosomes, and methods for delivery of the chromosomes to selected cells and tissues are provided. Also provided are cell lines for use in the methods, and cell lines and chromosomes produced by the methods. Methods for use of the artificial chromosomes are also provided.

Abstract: Production of proteins not normally secreted through conventional pathways such as membrane proteins including, for example, CFTR associated with cystic fibrosis, is now made possible by collection of such protein from the milk of lactating transgenic animals.

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: An isolated polypeptide comprising an amino acid sequence having at least 80% sequence identity to one or both of SEQ ID NOS:2 or 4, polynucleotides encoding these polypeptides, and antibodies to the polypeptides are useful in treating cancers.

Abstract: The invention is based on the discovery that certain variants of &bgr;-casein may induce Type-1 diabetes in susceptible individuals while other variants do not. The invention consists of the selection of non-diabetogenic milk producing cows and recovering and processing their milk and milk products. Another aspect of the invention is selectively breeding cows which produce the non-diabetogenic milk.

Abstract: The invention features methods of making transgenic animals, and transgenic animals made by such methods. The method includes introducing into a cell, a nucleic acid construct comprising a nucleic acid sequence encoding a heterologous polypeptide under the control of a mammary epithelial cell promoter, an insulator positioned 5′ from the promoter, an insulator positioned 3′ from the nucleic acid sequence encoding the polypeptide, and a prokaryotic sequence, wherein the sequence between the insulator 5′ from the promoter and the insulator 3′ from the nucleic acid encoding the polypeptide is substantially free of prokaryotic sequence; and allowing a transgenic mammal to develop from the cell, to thereby provide a transgenic mammal.

Abstract: A milk composition that includes a heterologous non-milk allergen is administered to a subject to suppress allergen-specific IgE production in the subject

Abstract: The invention provides modified prion-encoding genes for the creation of transgenic bovine and cervid animals resistant to transmissible spongiform encephalopathies including bovine spongiform encephalopathy (BSE). The transgenic animals homozygous for the mutant genes continue to express a functional copy of the prion-encoding gene, thereby not interfering with the normal role of the polypeptide and effectively decreasing tendency for alteration of sleep-wake cycles.