Abstract: The present invention provides modified oocytes having a nuclear genome derived from a first oocyte and cytoplasm derived from a second oocyte from a different subject, and methods for making and using such modified oocytes. The methods and compositions of the present invention can be useful in a variety of settings including, but not limited to, in in vitro fertilization (“IVF”) procedures.

Abstract: The present invention provides transgenic, large non-human animal models of cancer, as well as methods of using such animal models in the identification and characterization of therapies for cancer.

Abstract: The present invention provides modified oocytes having a nuclear genome derived from a first oocyte and cytoplasm derived from a second oocyte from a different subject, and methods for making and using such modified oocytes. The methods and compositions of the present invention can be useful in a variety of settings including, but not limited to, in in vitro fertilization (“IVF”) procedures.

Abstract: Methods are provided for producing a primate oocyte in vitro. The methods include removing nuclear DNA from a recipient primate oocyte from a first primate in a manner that does not lower levels of maturation promoting factor (MPF) to form an enucleated recipient primate oocyte. The recipient primate oocyte is enucleated using a non-UV-based spindle imaging system. Nuclear genetic material or DNA including chromosomes from a donor primate oocyte arrested at metaphase II from a second primate is isolated in the form of the karyoplast and introduced into the enucleated recipient primate oocyte. Introduction of the chromosomes is performed using a fusogenic agent or electroporation to produce a hybrid oocyte.

Abstract: A method of producing a chimera animal, characterized by transplanting, into a host animal, an aggregate of a first pluripotent stem cell that is of interest to strain establishment, a second pluripotent stem cell in which the ability to form germ cells is absent or decreased, and embryo.

Abstract: The present invention relates to a method for producing a transgenic pig in which immune rejection response is inhibited, and in which human HO-1 genes and TNFR1-Fe fusion genes are simultaneously expressed. The present invention also relates to a transgenic pig for organ transplantation, which is produced by the method, and in which immune rejection response is inhibited. The present invention also relates to a somatic-cell-donating cell strain for producing the transgenic pig, and to a method for producing organs, from the transgenic pig, in which the immune rejection response is inhibited.

Abstract: Methods and compositions are described for making phenotypically female fertile animals from XY donor cells and suitable host embryos. Culture media and methods are provided for maintaining XY donor cells in culture that after introduction into a host embryo and gestation in a suitable host will result in fertile XY female animals. Methods and compositions are described for making fertile female animals in an F0 generation from a donor XY cell and a host embryo, as are methods for making F1 progeny that are homozygous for a modification from a heterozygous F0 fertile male and a heterozygous F0 fertile female sibling.

Abstract: It is revealed that an organ such as pancreas can be regenerated by utilizing a fact that the deficiency of an organ is complemented by injecting an induced pluripotent stem cell (iPS cell) into a developed blastocyst in a blastocyst complementation method. Thus, the present invention has solved the above-described object. This provides a method for producing a target organ, using an iPS cell, in a living body of a non-human mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a different individual mammal that is an individual different from the non-human mammal.

Abstract: A composition and method for in vitro fertilization is provided which uses culture media comprising elevated concentrations of lipoic acid. More specifically, the invention provides culture media for developmental cells having a lipoic acid concentration of 5 ?M to 40 ?M. Culture media that include lipoic acid at concentrations within the identified range are able to provide blastocysts with increased survival, increased cell numbers, increased inner cell masses and/or increased percentage of the total mass made up by the inner cell compared to blastocysts cultured in a control medium.

Abstract: Methods of generating modified embryos and mammals by introduction of donor cells into an early stage embryo are provided, such that the resulting embryo and animal generated therefrom has a significant contribution to all tissues from the donor cells and is capable of transmitting the donor cell DNA.

Abstract: An embryonic stem cell line derived from a nucleus-transferred oocyte prepared by transferring a nucleus of a human somatic cell into an enucleated human oocyte may differentiate into various desired cell types.

Abstract: The present invention relates to a genetically modified pig as a model for studying atherosclerosis. The modified pig model displays one or more phenotypes associated with atherosclerosis. Disclosed is also a modified pig comprising a mutation in the endogenous ApoE gene or part thereof, LDL gene or part thereof, LDL receptor gene, or transcriptional or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of atherosclerosis; methods for screening the efficacy of a pharmaceutical composition; and a method for treatment of a human being suffering from atherosclerosis are disclosed.

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: The present invention discloses methods for cell nuclear transfer that comprise for example modification of zona pellucida of an oocyte, and/or sectioning of oocytes into several parts. The present invention also discloses methods for producing a genetically modified non-human mammal. Genetically modified non-human mammals obtainable by the disclosed methods are also within the scope of the present invention. Disclosed are also methods for cryopreservation of cells.

Abstract: The present invention relates to a modified pig as a model for studying Alzheimer's disease. The modified pig model displays one or more phenotypes associated with Alzheimer's disease. Disclosed is also a modified pig comprising a modified human and/or porcine APP gene, and/or PS1 gene, and/or a transcriptional and/or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of Alzheimer's disease; methods for screening the efficacy of a pharmaceutical composition; and a method for treatment of a human being suffering from Alzheimer's disease are disclosed.

Abstract: Use of PON gene cluster in preparing medicament for treating atherosclerosis in mammals, wherein the PON gene cluster treat atherosclerosis by promoting stability of atherosclerotic plaque. Method for the developing PON gene cluster transgenic mouse model and use of PON gene cluster in the development of PON gene cluster positive transgenic mouse model with atherosclerosis are also provided.

Abstract: An object of the invention is to effectively prepare a fish embryo with a correct chromosomal ploidy by nuclear transplantation in which an exogenous fish nucleus is transplanted in a cytoplasmic recipient. For this object, the invention comprises a step of preparing a fish embryo by transplanting a fish cell nucleus to an unfertilized egg. The step of preparing a fish embryo comprises a step of imposing physical and/or chemical stress to the unfertilized egg after activation. By imposing such stress, the stage of haplosis in a female nucleus which happens at the early stage of a series of developmental steps occurring in an unfertilized egg is suppressed and the correct ploidy of an obtained embryo is at least secured.

Abstract: Compositions and methods are provided for the efficient and reproducible generation of clone animals of all developmental stages. Also provided are methods of use of the same in reproductive and therapeutic cloning protocols.

Abstract: Methods and compositions for producing selected non-human mammalian germ cells and gametes and for making non-human animals using the produced germ cells and gametes are provided by the present invention. Methods of generating a non-human embryo and/or animal derived from donor stem cells, methods of generating chimeric non-human animals having substantially all gametes and/or germ cells derived from the donor stem cells, methods of producing a non-human host embryo lacking functional endogenous germ cells and non-human host embryos incapable of developing endogenous gametes of the present invention are described herein.

Abstract: Provided is a method for producing a cloned dog by enucleating an oocyte of a dog to produce an enucleated oocyte, transferring a somatic cell of the dog into the enucleated oocyte, carrying out electrofusion under optimized conditions to produce a nuclear transfer embryo, and transferring the nuclear transfer embryo into its surrogate mother.

Abstract: The present invention relates to a method for recombinant manufacturing of substances, wherein cells are transformed suing a nucleotide sequence coding for the substance, the transformed cells are subjected to a cloning process, and the cells obtained in this fashion are introduced into a host organism. The present invention relates in particular to the use of the method in the production of recombinant proteins, cells and tissues. According to a further aspect, the invention relates to a method, wherein the cells of an individual are isolated, said sells are introduced into an immunoincompetent animal for further growth and the cells, tissue and/or organs cultivated in the animal are again isolated and introduced into an individual.

Abstract: The present invention relates to a bovine beta-casein gene targeting vector comprising (1) a first region having a length of 5 to 12 kb which is homologous to the promoter and its flanking nucleic acid sequences of bovine beta-casein gene, and comprising exon 1, intron 1, and exon 2 of bovine beta-casein gene; (2) a region for cloning a nucleic acid coding for desired proteins; (3) a region for coding a positive selection marker; (4) a second region having a length of 2.8 to 3.5 kb which is homologous to the nucleic acid sequences of bovine beta-casein gene, and comprising exon 5, 6, 7 and 8, and intron 5, 6 and 7 of bovine beta-casein gene; wherein the nucleic acid segment corresponding to the first region is located upstream to the nucleic acid segment corresponding to the second region in the 5?-3? arrangement of beta-casein gene.

Abstract: This invention provides methods for producing antibodies, wherein the methods comprise the step of administering an immunogen comprising both a target antigen and a background antigen to transgenic animals, into which a gene coding for the background antigen has been introduced. Since immunotolerance to the background antigens have thus been induced in the transgenic animals, the animals efficiently produce antibodies to target antigens.

Abstract: The genome of the non-human mutant mammal, deficient in an endogenous Sigma receptor, contains a mutation that comprises a disruption in an endogenous Sigma receptor gene, wherein said gene disruption gives rise to a mutant lacking detectable levels of endogenous Sigma receptor. The mutant may be used as a control animal for in vivo tests, as well as a source of cells that can be used in in vitro tests. Mutants deficient in the Sigma-1 receptor can be used as models for in vivo study of disorders of the central nervous system, memory alterations, stress conditions and drug addictions, analgesia processes and neuroprotection. Mutants deficient in the Sigma-2 receptor can be used to study diagnostic or therapeutic tools to fight cancer and/or degenerative processes and/or to design compounds capable of preventing, reducing or alleviating the secondary pathology associated with administration of neuroleptic agents.

Abstract: The invention provides transgenic animals comprising a disruption in the endogenous GPR101 gene and methods of producing such transgenic animals. The invention further provides methods of identifying compounds that modulate GPR101 receptor proteins.

Abstract: Purified totipotent stem cells and pluripotent stems cells derived by somatic cell nuclear transfer are disclosed herein, as well as cell lines, multipotent cells and differentiated cells produced from these stem cells. The stem cells are produced from an enucleated host cell from a first donor and nuclear genetic material from a somatic cell of a second donor. Methods for making and using such compositions of such stem cells are also provided.

Abstract: The present invention relates, in general, to a method for introducing a heterologous replacement gene sequence into a host embryonic stem cell to replace an endogenous host gene target sequence. In particular, the invention relates to a method for inserting large pieces of DNA into embryonic stem cells with improved efficiency, by first deleting the endogenous host gene target sequence, and subsequently utilising two proximally positioned site-specific recombinase target (RT) sites to insert a heterologous replacement gene sequence into the host chromosome.

Abstract: The invention concerns the reconstruction in vitro of non-human mammal embryos by a method which consists in treating the nucleus of a somatic donor cell prior to its transfer into a receiver cytoplasm, said treatment comprising controlled proteolysis of non-histone proteins, and inducing an isomorphic swelling of said nucleus.

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: The present invention is related to a method for inducing in vitro the differentiation of stem cells or somatic cells towards germinal cells, comprising the following steps: cultivating said stem cells or somatic cells in a medium allowing their differentiation, and collecting the germinal cells from the culture medium, wherein said cells are cells transformed with an exogenous genetic construction comprising at least a Vasa gene. Germinal cells such as obtained and chimeric animals are also an object of this invention.

Abstract: The invention concerns a method for producing non-human mammal embryos, in particular rabbit by nuclear cloning. The invention also concerns the mammals obtained and their uses.

Abstract: An improved method of nuclear transfer employing long-term cultured somatic cells as the donor cells and enucleated oocytes as the recipient cells to produce dividing cybrids. Such cybrids are useful for developing viable animals clones when nurtured in a suitable host environment.

Abstract: Disclosed herein are a cloned canine and a production method thereof. The method comprises the steps of enucleating the oocyte of a canine to prepare an enucleated recipient oocyte, conducting nuclear transfer into the enucleated oocyte using a canine somatic cell as a nuclear donor cell under optimized conditions so as to prepare a nuclear transfer embryo, and transferring the nuclear transfer embryo into the oviduct of a surrogate mother. The present invention provides a method for producing cloned canines and thus, can contribute to the development of studies in veterinary medicine, anthropology and medical science such as the propagation of superior canines, the conservation of rare or nearly extinct canines, xenotransplantation and disease animal models.

Abstract: Disclosed herein is a method for producing a cyst-expressed transgenic animal using a PDK2 gene. The production method comprises preparing a PKD2 protein expression vector, inserting the expression vector into the nucleus of a fertilized egg to produce a PKD2 expression vector-containing fertilized egg, and transplanting the produced fertilized egg into the uterus of a surrogate mother. According to the invention disclosed herein, there is provided a method for producing transgenic animals, in which cysts are expressed only by the overexpression of the PKD2 gene. Also, transgenic mice are provided which can be effectively used in the investigation of cyst expression mechanisms and cyst control systems.

Abstract: This invention relates to a method for humanising a mouse. In particular, the invention relates to a method for replacing a cluster of mouse genes with single or multiple genes from the corresponding human cluster using a combination of homologous recombination and site-specific recombination.

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 provides novel transgenic nonhuman mammals capable of producing human sequence antibodies, as well as methods of producing and using these antibodies.

Abstract: The invention provides cloned transgenic ungulates (e.g., bovines) in which prion protein activity is reduced by one or more genetically engineered mutations. Desirably, these transgenic bovines are also genetically modified to express xenogenous (e.g., human) antibodies. Because of their resistance to prion-related diseases such as bovine spongiform encephalopy (also known as mad cow disease), these bovines are a safer source of human antibodies for pharmaceutical uses and a safer source of agricultural products.

Abstract: The invention relates to a method of producing a protein of interest, comprising making a non-human transgenic mammal that produces said protein in its milk, obtaining said milk from the non-human transgenic mammal and purifying said protein of interest from the milk. Transgenic bovine animals were generated, which are able to produce human growth hormone in mammary glands. The method involves cloning of a genetic construct encoding hGH gene and beta casein promoter conveniently in an expression vector. It also includes transfection procedures into fetal bovine somatic cells, generally fibroblasts, and the nuclear transfer into enucleated bovine oocytes, generating thus transgenic embryos.

Abstract: The present invention is directed to various methodologies to make NT a practical procedure for animals, specifically, primates including human and non-human primates. Furthermore, the methods and molecular components provided by the present invention provide a practical means for producing embryos with desired characteristics. In a specific embodiment, the methodology of the present invention comprises introducing nuclei having desired characteristics along with one or more molecular components into an enucleated egg, thus creating a nuclear transfer construct, culturing the egg to produce a viable embryo, transferring the embryo to the oviducts of a female, and producing a cloned animal.

Abstract: The invention relates to a method for cloning in the rat by nuclear transfer. The invention further relates to the rats obtained thus, in the foetal or adult state, as well as use thereof for the production of molecules of interest or as study models.

Abstract: Method of culturing embryonic stem (ES) cells of avian origin includes the steps of: a) suspending ES cells originating from the blastoderm disk of fertilized un-incubated avian egg(s) in a basal culture medium supplemented with: insulin-like growth factor-1 (IGF-1) and ciliary neurotrophic factor (CNTF); and animal serum; and, optionally, at least one growth factor selected from among interleukin 6 (II-6), interleukin 6 receptor (II-6R), stem cell factor (SCF), fibroblast growth factor (FGF), leukemia inhibitory factor (LIF), interleukin 11 (II-11), oncostatin and/or cardiotrophin; b) seeding the suspension of ES cells obtained in step a) on a layer of feeder cells and further culturing the ES cells for at least 2 to 10 passages; c) optionally, removing at least one growth factor selected from among SCF, FGF, II-6, II-6R, LIF, oncostatin, cardiotrophin and II-11 from the culture medium; and d) further culturing the ES cells in the medium of step c) on a layer of feeder cells.

Abstract: The present invention is a porcine animal, tissue, organ, cells and cell lines, which lack any expression of functional alpha 1,3 galactosyltransferase (alpha1,3GT). These animals, tissues, organs and cells can be used in xenotransplantation and for other medical purposes.

Abstract: Disclosed herein are methods and compositions for genome editing of one or more loci in a rat, using fusion proteins comprising a zinc-finger protein and a cleavage domain or cleavage half-domain. Polynucleotides encoding said fusion proteins are also provided, as are cells comprising said polynucleotides and fusion proteins.

Abstract: Compositions and methods are provided for the efficient and reproducible generation of clone animals of all developmental stages. Also provided are methods of use of the same in reproductive and therapeutic cloning protocols.

Abstract: The invention pertains to the discovery that the presence of oocyte spindle associated factors in an enucleated oocyte improves oocyte quality and subsequently nuclear transfer. In particular, it was discovered that maintaining oocyte spindle factors in the oocyte after enucleation improves oocyte quality for use in nuclear transfer methodology.

Abstract: The present invention relates to a genetically modified pig comprising at least one site for integration of at least one transgene. The invention also pertains to a porcine embryo, blastocyst, foetus, donor cell and/or cell nucleus, derived from said genetically modified pig. In another aspect, the invention relates to any genetically modified porcine blastocyst, wherein the genetically modified genome comprises at least one site for integration of at least one transgene.

Abstract: The present invention relates to a modified pig as a model for studying Alzheimer's disease. The modified pig model displays one or more phenotypes associated with Alzheimer's disease. Disclosed is also a modified pig comprising a modified human and/or porcine APP gene, and/or PS1 gene, and/or a transcriptional and/or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of Alzheimer's disease; methods for screening the efficacy of a pharmaceutical composition; and a method for treatment of a human being suffering from Alzheimer's disease are disclosed.

Abstract: The invention relates to a method of producing a protein of interest, comprising making a non-human transgenic mammal that produces said protein in its milk, obtaining said milk from the non-human transgenic mammal and purifying said protein of interest from the milk. Transgenic bovine animals were generated, which are able to produce human growth hormone in mammary glands. The method involves cloning of a genetic construct encoding hGH gene and beta casein promoter conveniently in an expression vector. It also includes transfection procedures into fetal bovine somatic cells, generally fibroblasts, and the nuclear transfer into enucleated bovine oocytes, generating thus transgenic embryos.

Abstract: The present invention relates to a genetically modified pig as a model for studying psoriasis. The modified pig model displays one or more phenotypes associated with psoriasis. Disclosed is also a modified pig comprising a mutation in the endogenous ILK-I Ra, JunB/cJun, CD18, IKK2, and/or LIG1 gene, and/or a human, porcine and/or murine PPARs, PPAR-?, l?B-?, STAT3c, Integrin beta 1, Integrin alpha 2, MEK1, Amphiregulin, BMP-6, VEGF, JunB?ec-Jun?ep, IL-I a, TGF.beta 1, CD18 hypo, Cre/lkk2FL/FL, Dsg1, SCCE, TGF-a, TNF-a, IL-20, IFN-g, LIG1 KO, KGF, IL-6, PAFR1 Cre/lkk2FL/FL, IL1 R, Dsg3, IFN-gamma, p40, ILI Ra, IKK2, JunB/c-Jun, and/or LIG1 gene, transcriptional and/or translational product or part thereof. The invention further relates to methods for producing the modified pig; and methods for evaluating the effect of a therapeutical treatment of psoriasis, for screening the efficacy of a pharmaceutical composition, and a method for treatment of human being suffering from psoriasis are disclosed.