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1. 发明授权

US06331659B1 Cumulus cells as nuclear donors 有权
标题翻译：积细胞作为核供体
公开(公告)号：US06331659B1
公开(公告)日：2001-12-18
申请号：US09132104
申请日：1998-08-10
申请人： Teruhiko Wakayama , Ryuzo Yanagimachi
发明人： Teruhiko Wakayama , Ryuzo Yanagimachi
IPC分类号： C12N1500
CPC分类号： C12N15/8775
摘要： Animals are produced following injection of adult somatic cell nuclei into enucleated oocytes. The invention provides a method for cloning an animal by directly inserting at least a portion of the adult somatic nucleus (including the minimum chromosomal material able to support development) into a recipient enucleated oocyte. Preferably, the nucleus is inserted by microinjection and, more preferably, by piezo electrically-actuated microinjection. The oocyte is activated prior to, during, or up to about 6 hours after insertion of the nucleus, by electroactivation or exposure to a chemical activating agent, such as Sr2+. The activated renucleated oocyte is allowed to develop into an embryo and is transplanted to a host surrogate mother to develop into a live offspring.
摘要翻译：在将成体体细胞核注射到去核卵母细胞后产生动物。本发明提供了一种用于通过将至少一部分成体体细胞核（包括能够支持发育的最小染色体材料）直接插入受体去核卵母细胞来克隆动物的方法。优选地，通过显微注射插入细胞核，更优选地，通过压电电致动显微注射。卵母细胞在插入核后之前，期间或至多约6小时期间被激活，通过电活化或暴露于化学活化剂如Sr2 +。将激活的复核卵母细胞发育成胚胎，并移植到宿主代孕母体中以形成活的后代。

2. 发明授权

US06641526B1 Development of normal offspring from oocytes injected with freeze-dried spermatozoa 有权
标题翻译：从注射冷冻干燥精子的卵母细胞培养正常后代
公开(公告)号：US06641526B1
公开(公告)日：2003-11-04
申请号：US09177391
申请日：1998-10-23
申请人： Teruhiko Wakayama , Ryuzo Yanagimachi
发明人： Teruhiko Wakayama , Ryuzo Yanagimachi
IPC分类号： A61B1743
CPC分类号： A01N1/02 , A01N1/0284
摘要： The invention provides a method for freeze-drying spermatozoa to obtain at least one reconstituted spermatozoon whose head (nucleus) is capable of fertilizing an oocyte to produce a live offspring. The motility of spermatozoa which have been freeze-dried and stored in a vacuum at room temperature is not restored when rehydrated. Their plasma membranes are disrupted and they are all “dead” in the conventional sense. However, when they are injected microsurgically into oocytes, their nuclei transform into male pronuclei and participate in normal embryonic development.
摘要翻译：本发明提供了一种用于冷冻精子的方法，以获得至少一个重构的精子，其头（核）能够施用卵母细胞以产生活的后代。冷冻干燥并在室温真空中储存的精子的运动性在再水合时不会恢复。他们的等离子体膜被破坏，它们在常规意义上都是“死的”。然而，当它们被显微外科注射到卵母细胞中时，它们的核转变成雄性原核并参与正常的胚胎发育。

3. 发明授权

US6143564A Use of the polar body chromosomes for the production of embryos and normal offspring 失效
标题翻译：使用极体染色体来生产胚胎和正常的后代
公开(公告)号：US6143564A
公开(公告)日：2000-11-07
申请号：US227365
申请日：1999-01-08
申请人： Teruhiko Wakayama , Ryuzo Yanagimachi
发明人： Teruhiko Wakayama , Ryuzo Yanagimachi
IPC分类号： C12N15/877 , C12N15/00 , A01K67/00 , A01K67/027 , C12N15/02 , C12N15/63
CPC分类号： C12N15/8775
摘要： The invention provides a method for obtaining a live offspring having maternal chromosomes derived from a live first polar body of an oocyte that has completed the first meiotic division. It has been discovered herein that chromosomes in the first polar body are able to participate in normal embryonic development if they are allowed to complete the second meiotic division within an enucleated mature oocyte, and are then allowed to mingle with chromosomes of a spermatozoon. The invention further provides a method for producing up to four embyros or live offspring having the chromosomes of a single oocyte, by using both the first polar body chromosomes, and the second polar body chromosomes to reconstitute recipient enucleated (fertilized) oocytes.
摘要翻译：本发明提供了一种获得具有源自卵母细胞的活的第一极体的母体染色体的活后代的方法，其完成了第一次减数分裂。在本文中已经发现，如果允许其在去核的成熟卵母细胞内完成第二次减数分裂，则第一极体中的染色体能够参与正常的胚胎发育，然后允许其与精子的染色体相互融合。本发明还提供了通过使用第一极体染色体和第二极体染色体来重构受体去核（受精）卵母细胞来生产多达四个具有单个卵母细胞染色体的雌雄或活后代的方法。

4. 发明授权

US07138227B2 Frozen spermatozoa compositions and uses thereof 失效
公开(公告)号：US07138227B2
公开(公告)日：2006-11-21
申请号：US10259553
申请日：2002-09-27
申请人： Hirokazu Kusakabe , Monika Anna Sczcygiel , David Gordon Whittingham , Ryuzo Yanagimachi , Takehito Kaneko
发明人： Hirokazu Kusakabe , Monika Anna Sczcygiel , David Gordon Whittingham , Ryuzo Yanagimachi , Takehito Kaneko
IPC分类号： A61K35/52
CPC分类号： A01N1/02 , A01N1/0221
摘要： The present invention provides a composition for freezing or freeze-drying spermatozoa or sperm heads thereof, wherein the composition enables the spermatozoa or sperm heads thereof to maintain chromosome integrity at a temperature of about +4° C. to about −200° C. Also provided is a container containing the composition. The present invention further provides a method for maintaining chromosome integrity of spermatozoa or sperm heads during freezing or freeze-drying. The present invention also provides methods for freezing or freeze-drying spermatozoa to obtain at least one spermatozoan capable of fertilizing an oocyte to produce a live offspring. Also provided are frozen or freeze-dried spermatozoa produced by these methods, and containers containing the frozen or freeze-dried spermatozoa. Finally, the present invention provides methods for producing a live mammalian offspring from an oocyte fertilized with a rehydrated freeze-dried (or thawed or freeze-thawed) spermatozoan.

5. 发明授权

US06376743B1 Mammalian transgenesis by intracytoplasmic sperm injection 有权
标题翻译：通过胞质内精子注射的哺乳动物转基因
公开(公告)号：US06376743B1
公开(公告)日：2002-04-23
申请号：US09371648
申请日：1999-08-10
申请人： Ryuzo Yanagimachi
发明人： Ryuzo Yanagimachi
IPC分类号： A01K67027
CPC分类号： A01K67/0275 , A01K67/0335 , A01K67/0338 , A01K2217/05 , C12N15/87 , C12N15/89
摘要： Coinjection of unfertilized mouse oocytes with sperm heads and exogenous nucleic acid encoding a transgene results in transgene-expressing embryos, reflecting nucleic acid-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos resulting from coinjection produced offspring expressing the integrated transgene.
摘要翻译：注射未受精的小鼠卵母细胞与精子头和编码转基因的外源核酸导致转基因表达的胚胎，反映核心 - 精子头关联在共注射之前。转移到由共同产生的胚胎替代母亲的无效转移产生表达整合的转基因的后代。

6. 发明申请

US20060294606A1 Tn5 transposase-mediated transgenesis 审中-公开
标题翻译： Tn5转座酶介导的转基因
公开(公告)号：US20060294606A1
公开(公告)日：2006-12-28
申请号：US11127685
申请日：2005-05-11
申请人： Istefo Moisyadi , Ryuzo Yanagimachi , Ryota Suganuma
发明人： Istefo Moisyadi , Ryuzo Yanagimachi , Ryota Suganuma
IPC分类号： A01K67/027
CPC分类号： C12N15/90 , A01K67/0275 , A01K2217/05 , A01K2227/105 , C12N9/22
摘要： The present invention relates to methods for obtaining transgenic embryos, animals, and plants. Also encompassed in the present invention are transposase-mediated trangenesis methods including transposase-mediated intracytoplasmic sperm injection (TN:ICSI), transposase-mediated intracytoplasmic round spermatid injection (TN:ROSI), and transposase-mediated in vitro fertilization (TN:IVF).
摘要翻译：本发明涉及获得转基因胚胎，动物和植物的方法。本发明还包括转座酶介导的转移方法，包括转座酶介导的胞质内精子注射（TN：ICSI），转座酶介导的胞质内圆形精子细胞注射（TN：ROSI）和转座酶介导的体外受精（TN：IVF）。

7. 发明申请

US20060031952A1 Methods for producing transgenic animals 审中-公开
标题翻译：生产转基因动物的方法
公开(公告)号：US20060031952A1
公开(公告)日：2006-02-09
申请号：US11151936
申请日：2005-06-14
申请人： Pawel Pelczar , Stefan Moisyadi , Barbara Hohn , Takehito Kaneko , Ryuzo Yanagimachi
发明人： Pawel Pelczar , Stefan Moisyadi , Barbara Hohn , Takehito Kaneko , Ryuzo Yanagimachi
IPC分类号： A01K67/027 , C12N5/08 , C12N5/06
CPC分类号： C12N15/907 , A01K67/0275 , A01K2217/05 , C12N2517/10
摘要： The invention provides methods for producing transgenic embryos and animals with a higher efficiency than presently available procedures based on Intracytoplasmic Sperm Injection (ICSI) of nucleic acid complexed with recombinase.
摘要翻译：本发明提供了基于与重组酶复合的核酸的胞质内精子注射（ICSI）相比，生产具有比目前可用的程序更高效率的转基因胚胎和动物的方法。

8. 发明授权

US06593139B1 Initiation of full mammalian oocyte activation by multiple sperm components 失效
标题翻译：通过多种精子成分启动完全哺乳动物卵母细胞活化
公开(公告)号：US06593139B1
公开(公告)日：2003-07-15
申请号：US09478848
申请日：2000-01-07
申请人： Ryuzo Yanagimachi , Anthony C. F. Perry
发明人： Ryuzo Yanagimachi , Anthony C. F. Perry
IPC分类号： C12N500
CPC分类号： C12N5/0609 , C07K14/4705 , C12N2500/84 , C12N2517/10
摘要： The invention provides a microinsertion-based, trans-complementation method of in vitro oocyte activation useful to identify properties of sperm-borne oocyte-activating factor(s) (SOAF) and ooplasmic interactions with sperm components at fertilization. The invention provides at least one detergent-insoluble heat-sensitive component of the perinuclear matrix of a spermatozoon (SOAFm) that acts coordinately with at least one heat-stable submembrane sperm component. SOAFm is solubilized in vitro (SOAFs) under reducing conditions similar to those encountered in the ooplasm. By the method of the invention, the failure of heat-inactivated demembranated sperm heads to activate an egg is rescued by coinsertion of SOAFs into an oocyte. SOAFs is protease-sensitive and is liberated from demembranated heads in a temperature-dependent manner that inversely correlates with the ability of demembranated sperm heads to activate oocytes.
摘要翻译：本发明提供了体外卵母细胞激活的基于微插入的反互补方法，其用于鉴定精子传播的卵母细胞活化因子（SOAF）的性质以及在受精时与精子成分的卵泡相互作用。本发明提供了至少一种与至少一种热稳定的亚膜精子成分协同作用的精子（SOAFm）的核周基质的至少一种不溶于洗涤剂的热敏成分。 SOAFm在与卵泡中遇到的相似的还原条件下在体外溶解（SOAF）。通过本发明的方法，通过将SOAFs插入卵母细胞而获得热灭活的精子头激活卵的失败。 SOAF是蛋白酶敏感的，并且以温度依赖的方式从分化的头部释放，与分化的精子头激活卵母细胞的能力成反比关系。

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