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31. 发明申请

US20060064763A1 Unactivated oocytes as cytoplast recipients for nuclear transfer 失效
公开(公告)号：US20060064763A1
公开(公告)日：2006-03-23
申请号：US11265114
申请日：2005-11-03
申请人： Keith Campbell , Ian Wilmut
发明人： Keith Campbell , Ian Wilmut
IPC分类号： A01K67/027
CPC分类号： C12N15/8773 , A61K35/54 , C12N15/873 , C12N15/8771
摘要： 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.

32. 发明申请

US20060026694A1 Method for generating immune-compatible cells and tissues using nuclear transfer techniques 审中-公开
公开(公告)号：US20060026694A1
公开(公告)日：2006-02-02
申请号：US11130979
申请日：2005-05-17
申请人： Robert Lanza , Michael West
发明人： Robert Lanza , Michael West
IPC分类号： A01K67/027
CPC分类号： C12N15/873 , A01K67/0271 , A01K2217/05 , A01K2227/101 , A01K2227/105 , A01K2267/025 , A61K49/0008 , C12N15/8771 , C12N2517/04
摘要： This invention relates to methods for making immune compatible tissues and cells for the purpose of transplantation and tissue engineering, using the techniques of nuclear transfer and cloning. Also encompassed are methods for determining the effect on immune compatibility of expressed transgenes and other genetic manipulations of the engineered cells and tissues.

33. 发明申请

US20060021070A1 Production of chimeric bovine or porcine animals using cultured inner cell mass 审中-公开
标题翻译：使用培养的内细胞块生产嵌合牛或猪动物
公开(公告)号：US20060021070A1
公开(公告)日：2006-01-26
申请号：US11024010
申请日：2004-12-28
申请人： Steven Stice , Jose Cibelli , James Robl , Paul Golueke , F. Leon , D. Jerry
发明人： Steven Stice , Jose Cibelli , James Robl , Paul Golueke , F. Leon , D. Jerry
IPC分类号： A01K67/027
CPC分类号： C12N15/8771 , A01K2217/05 , C12N15/8778
摘要： Novel cultured inner cell mass (CICM) cells, and cell lines, derived from ungulates, in particular, pigs and cows, and methods for their preparation are provided. The subject CICMs possess similar morphology and express cell markers identically or substantially similarly to ICMs of undifferentiated developing embryos for prolonged culturing periods. Heterologous DNA is inserted into the subject CICM cells and cell lines so as produce transgenic CICM cell which are introduced into non-human fertilized embryos to produce transgenic chimeric embryos. The transgenic chimeric embryos are transferred into recipient females where they are permitted to develop into transgenic chimeric fetuses. Recipient females give birth to transgenic chimeric animals which are capable of transmitting the heterologous DNA to their progeny. Transgenic CICM cells are also used to produce cloned transgenic embryos, fetuses and offspring.
摘要翻译：提供了新颖的培养的内细胞团（CICM）细胞和源自有蹄类动物，特别是猪和牛的细胞系及其制备方法。受试者CICM具有与未分化发育的胚胎的ICM相似或基本上类似于延长培养期的相似形态和表达细胞标记。将异源DNA插入受试者CICM细胞和细胞系中，以产生转入非人受精胚胎以产生转基因嵌合胚胎的转基因CICM细胞。将转基因嵌合胚胎转移到受体雌性中，在其中允许其转化为转基因嵌合胎儿。受体女性产生能够将异源DNA转移到其后代的转基因嵌合动物。转基因CICM细胞也用于产生克隆的转基因胚胎，胎儿和后代。

34. 发明申请

US20050177879A1 Process for producing exogenous protein in the milk of transgenic mammals and a process for purifying proteins therefrom 有权
标题翻译：用于在转基因哺乳动物的乳中产生外源蛋白质的方法和从其中纯化蛋白质的方法
公开(公告)号：US20050177879A1
公开(公告)日：2005-08-11
申请号：US10952377
申请日：2004-09-29
申请人： Carlos Melo , Lino Baranao , Cesar Carbonetto
发明人： Carlos Melo , Lino Baranao , Cesar Carbonetto
IPC分类号： A01K67/027 , C07K1/00 , C07K14/00 , C07K14/61 , C07K16/00 , C07K17/00 , C12N20060101 , C12N15/00 , C12N15/85 , C12N15/877 , C12P21/00
CPC分类号： A01K67/0275 , A01K2217/05 , A01K2227/101 , A01K2267/01 , C07K14/61 , C12N15/8509 , C12N15/8771 , C12N2830/008
摘要： 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. The method also includes other procedures to generate transgenic embryos for the further expansion of the transgenic herd, such as the subcloning of transgenic female bovines, the superovulation of transgenic cows and their insemination with semen from a non-transgenic or a transgenic male bovine, and the superovulation of non-transgenic cows and their insemination with semen from a transgenic male bovine. Afterwards, transgenic embryos give rise to transgenic cattle that produce human growth hormone in huge amounts in their milk, from which the hormone is completely purified and analysed to fulfill all the requirements for the manufacture of a pure biopharmaceutical product.
摘要翻译：本发明涉及产生感兴趣的蛋白质的方法，包括制备在其乳中产生所述蛋白质的非人转基因哺乳动物，从非人转基因哺乳动物获得所述乳，并从所述牛奶中纯化所述目的蛋白质。产生转基因牛动物，其能够在乳腺中产生人生长激素。该方法包括在表达载体中方便地克隆编码hGH基因和β酪蛋白启动子的遗传构建体。它还包括将转染过程转化为胎牛体细胞，通常是成纤维细胞，以及将核转移到去核的牛卵母细胞中，从而产生转基因胚胎。该方法还包括产生用于进一步扩增转基因畜群的转基因胚胎的其他方法，例如转基因雌性牛的亚克隆，转基因奶牛的超排卵及其来自非转基因或转基因雄性牛的精液授精，以及非转基因奶牛的超排卵及其来自转基因雄性牛的精液授精。之后，转基因胚胎产生大量牛奶中产生人生长激素的转基因牛，激素被完全纯化和分析，以满足制造纯生物药物产品的所有要求。

35. 发明申请

US20040083498A1 Cloning method using oocytes matured in vivo 审中-公开
标题翻译：使用卵母细胞在体内成熟的克隆方法
公开(公告)号：US20040083498A1
公开(公告)日：2004-04-29
申请号：US10414458
申请日：2003-04-14
发明人： Paul Alexandre DeSousa , Timothy James King , Ian Wilmut , Jie Zhu
IPC分类号： A01K067/027
CPC分类号： C12N15/8771 , A61K38/24 , C12N5/0609 , C12N15/8778 , C12N2500/02 , C12N2501/31 , C12N2502/243 , C12N2517/10
摘要： A method is described of selecting a porcine oocyte or a population of oocytes with improved developmental competence following either in vitro or in vivo maturation.
摘要翻译：描述了在体外或体内成熟后选择具有改善的发育能力的猪卵母细胞或卵母细胞群的方法。

36. 发明申请

US20040068760A1 Transgenic ungulates capable of human antibody production 失效
标题翻译：能够产生人抗体的转基因有蹄类动物
公开(公告)号：US20040068760A1
公开(公告)日：2004-04-08
申请号：US10441503
申请日：2003-05-19
发明人： James M. Robl , Philippe Collas , Eddie Sullivan , P. Kasinathan , Richard A. Goldsby , Yoshimi Kuroiwa , Kazuma Tomizuka , Isao Ishida
IPC分类号： A01K067/027
CPC分类号： A01K67/0273 , A01K67/0275 , A01K67/0278 , A01K2207/15 , A01K2217/00 , A01K2267/01 , C07K16/00 , C07K16/42 , C07K2317/21 , C12N15/8509 , C12N15/8771 , C12N2799/025 , C12N2800/208
摘要： The invention features novel methods for the production of large quantities of xenogenous antibodies, such as human antibodies. Preferably, this result is effected by inactivation of IgM heavy chain expression and, optionally, by inactivation of Ig light chain expression, and by the further introduction of an artificial chromosome which results in the expression of xenogenous antibodies (e.g., non-bovine antibodies), preferably human antibodies.
摘要翻译：本发明的特征在于产生大量异种抗体（例如人抗体）的新方法。优选地，该结果通过IgM重链表达的失活和任选地通过Ig轻链表达的失活以及进一步引入导致异源抗体（例如非牛抗体）的表达的人造染色体来实现，，优选人抗体。

37. 发明申请

US20030037352A1 Unactivated oocytes as cytoplast recipients for nuclear transfer 审中-公开
公开(公告)号：US20030037352A1
公开(公告)日：2003-02-20
申请号：US09973701
申请日：2001-10-11
申请人： Roslin Institute (Edinburgh)
发明人： Keith Henry Stockman Campbell , Ian Wilmut
IPC分类号： A01K067/027
CPC分类号： C12N15/8773 , A61K35/54 , C12N15/873 , C12N15/8771
摘要： 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.

38. 发明授权

US06395958B1 Method of producing a polypeptide in an ungulate 有权
标题翻译：在有蹄类动物中产生多肽的方法
公开(公告)号：US06395958B1
公开(公告)日：2002-05-28
申请号：US09354276
申请日：1999-07-15
申请人： Nikolai S. Strelchenko , Jeffrey M. Betthauser , Gail L. Jurgella , Marvin M. Pace , Michael D. Bishop
发明人： Nikolai S. Strelchenko , Jeffrey M. Betthauser , Gail L. Jurgella , Marvin M. Pace , Michael D. Bishop
IPC分类号： C12P2100
CPC分类号： C12N15/8771 , C12N5/0603 , C12N5/0611 , C12N2502/13 , C12N2510/04
摘要： 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.
摘要翻译：本发明涉及克隆技术。本发明部分涉及用于克隆动物的永生化和全能细胞，使用核转移技术从这些细胞产生的胚胎，从这些细胞和胚胎产生的动物，以及用于建立这些细胞和胚胎的材料，方法和方法动物。

39. 发明申请

US20020035737A1 Cloning pigs using donor nuclei from differentiated cells 审中-公开
标题翻译：使用来自分化细胞的供体核克隆猪
公开(公告)号：US20020035737A1
公开(公告)日：2002-03-21
申请号：US09845353
申请日：2001-05-01
申请人： University of Massachusetts
发明人： Steven L. Stice , James M. Robl , Jose Cibelli , Paul Golueke
IPC分类号： A01K067/027
CPC分类号： C12N15/8778 , A01K67/027 , A01K67/0273 , A01K67/0275 , A01K2217/05 , A01K2227/101 , A61K35/12 , A61K38/00 , C12N5/0603 , C12N15/8771 , C12N2510/00 , C12N2517/02 , C12N2517/04
摘要： An improved method of nuclear transfer involving the transplantation of donor differentiated pig cell nuclei into enucleated pig oocytes is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring. Production of genetically engineered or transgenic pig embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
摘要翻译：提供了涉及将供体分化的猪细胞核移植到去核猪卵母细胞中的改进的核转移方法。所产生的核转移单位可用于通过胎儿和后代的产生来增殖基因型和转基因基因型。通过本发明的方法促进转基因植物或转基因猪胚胎，胎儿和后代的生产，因为供体核的分化细胞源可被遗传修饰和克隆繁殖。

40. 发明申请

US20020010949A1 Cloning using donor nuclei from differentiated fetal and adult cells 审中-公开
标题翻译：使用来自分化胎儿和成体细胞的供体核克隆
公开(公告)号：US20020010949A1
公开(公告)日：2002-01-24
申请号：US09828876
申请日：2001-04-10
申请人： University of Massachusetts, Amherst Campus
发明人： Steven L. Stice , Jose Cibelli , James Robl , Paul Golueke , F. Abel Ponce de Leon , D. Joseph Jerry
IPC分类号： A01K067/00 , A61K048/00
CPC分类号： A01K67/0275 , A01K67/027 , A01K2217/05 , A01K2227/101 , A01K2227/106 , A61K35/12 , C12N5/0603 , C12N15/8771 , C12N15/8778 , C12N2510/00 , C12N2517/02 , C12N2517/04
摘要： An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
摘要翻译：提供了涉及将供体分化的细胞核移植到与供体细胞相同物种的去核卵母细胞中的改进的核转移方法。所产生的核转移单位可用于通过胎儿和后代的产生以及用于产生包括人同种异体胚胎或干细胞在内的等基因CICM细胞的基因型和转基因基因型的倍增。通过本方法促进遗传工程或转基因哺乳动物胚胎，胎儿和后代的生产，因为供体核的分化细胞源可以被遗传修饰和克隆繁殖。

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