公开(公告)号：US08592643B2

公开(公告)日：2013-11-26

申请号：US12865304

申请日：2008-12-09

申请人： Erika Sasaki ,  Hideyuki Okano

发明人： Erika Sasaki ,  Hideyuki Okano

IPC分类号： A01K67/00

CPC分类号： A01K67/0275 ,  A01K2217/052 ,  A01K2227/106 ,  A01K2267/0318 ,  A01K2267/0356 ,  A01K2267/0393 ,  C07K14/43595 ,  C07K14/47 ,  C12N15/8509 ,  C12N2740/15043 ,  C12N2799/027 ,  C12N2999/007

摘要： An object of the present invention is to provide a method for introducing a gene into an embryo for production of a human disease model primate animal using a non-human primate animal such as a marmoset. The present invention relates to a method for introducing a foreign gene into an early embryo of a non-human primate animal, which comprises placing early embryos of a non-human primate in a 0.2 M to 0.3 M sucrose solution, so as to increase the volume of the perivitelline spaces, and then injecting a viral vector containing a human foreign gene operably linked to a promoter into the perivitelline spaces of the early embryos.

摘要翻译： 本发明的目的是提供一种使用非人灵长类动物如for子将基因引入胚胎以产生人类疾病模型灵长类动物的方法。 本发明涉及将外源基因导入非人灵长类动物的早期胚胎的方法，其包括将非人灵长类动物的早期胚胎置于0.2M至0.3M蔗糖溶液中，以增加 然后将包含可操作地连接到启动子的人外源基因的病毒载体注入早期胚胎的周围组织中。

公开(公告)号：US20130312127A1

公开(公告)日：2013-11-21

申请号：US13982418

申请日：2011-12-02

申请人： Ten-Tsao Wong ,  Paul Collodi

发明人： Ten-Tsao Wong ,  Paul Collodi

IPC分类号： C12N15/85

CPC分类号： C12N15/8509 ,  A01K67/0275 ,  A01K2217/052 ,  A01K2217/203 ,  A01K2217/206 ,  A01K2227/40 ,  A01K2267/02 ,  A01K2267/0393 ,  C07K14/522 ,  C07K14/5415

摘要： Methods of disrupting germ cell migration and development in a fish embryo by inducing targeted expression of Sdf-1 a or Lif and disruption of the Sdf-1 a gradient or a Lif signaling pathway in the fish embryo have been developed. Plasmid constructs containing a gene encoding Sdf-1 a or Lif and a gene encoding a signaling sequence for targeted expression of Sdf-1 a or Lif have been generated. The plasmids will be administered to a fish or a population of fish to reproductively sterilize the population with efficacy of up to 100%. Transgenic fish of this invention are reproductively incompetent of genetically contaminating a wild fish population.

摘要翻译： 已经开发出通过诱导Sdf-1α的靶向表达或者在鱼胚胎中产生Sdf-1梯度或Lif信号通路的Lif和破坏来破坏鱼胚胎中生殖细胞迁移和发育的方法。 已经产生了含有编码Sdf-1a或Lif的基因和编码用于靶向表达Sdf-1a或Lif的信号序列的基因的质粒构建体。 质粒将被给予鱼或鱼群，以高达100％的效力对人群进行再生灭菌。 本发明的转基因鱼在遗传上污染野生鱼群体的生殖能力不足。

公开(公告)号：US20130212725A1

公开(公告)日：2013-08-15

申请号：US13702231

申请日：2011-06-07

申请人： Ralf Kühn ,  Wolfgang Wurst ,  Melanie Meyer

发明人： Ralf Kühn ,  Wolfgang Wurst ,  Melanie Meyer

IPC分类号： C12N15/62

CPC分类号： C12N15/62 ,  A01K67/0276 ,  A01K67/0278 ,  A01K2207/05 ,  A01K2217/07 ,  A01K2217/072 ,  A01K2227/105 ,  A01K2267/03 ,  A01K2267/0393 ,  C07K2319/80 ,  C12N9/22 ,  C12N15/907

摘要： The present invention relates to a method of modifying a target sequence in the genome of a eukaryotic cell, the method comprising the step: (a) introducing into the cell a fusion protein comprising a DNA-binding domain of a Tal effector protein and a non-specific cleavage domain of a restriction nuclease or a nucleic acid molecule encoding the fusion protein in expressible form, wherein the fusion protein specifically binds within the target sequence and introduces a double strand break within the target sequence. The present invention further relates to the method of the invention, wherein the modification of the target sequence is by homologous recombination with a donor nucleic acid sequence further comprising the step: (b) introducing a nucleic acid molecule into the cell, wherein the nucleic acid molecule comprises the donor nucleic acid sequence and regions homologous to the target sequence. The present invention also relates to a method of producing a non-human mammal or vertebrate carrying a modified target sequence in its genome. Furthermore, the present invention relates to a fusion protein comprising a DNA-binding domain of a Tal effector protein and a non-specific cleavage domain of a restriction nuclease.

摘要翻译： 本发明涉及一种修饰真核细胞基因组中的靶序列的方法，所述方法包括以下步骤：（a）将包含Tal效应蛋白的DNA结合结构域的融合蛋白引入细胞， 限制性核酸酶或编码可表达形式的融合蛋白的核酸分子的特异性切割结构域，其中所述融合蛋白在靶序列内特异性结合并在靶序列内引入双链断裂。 本发明还涉及本发明的方法，其中靶序列的修饰是通过与供体核酸序列的同源重组而进一步包括步骤：（b）将核酸分子引入细胞，其中核酸 分子包含供体核酸序列和与靶序列同源的区域。 本发明还涉及在其基因组中产生携带修饰的靶序列的非人哺乳动物或脊椎动物的方法。 此外，本发明涉及包含Tal效应蛋白的DNA结合结构域和限制性核酸酶的非特异性切割结构域的融合蛋白。

公开(公告)号：US20130198874A1

公开(公告)日：2013-08-01

申请号：US13581510

申请日：2011-03-04

申请人： Masaru Okabe ,  Masahito Ikawa ,  Tadashi Kimura ,  Keiichi Kumasawa

发明人： Masaru Okabe ,  Masahito Ikawa ,  Tadashi Kimura ,  Keiichi Kumasawa

IPC分类号： A01K67/027 ,  A61D19/04

CPC分类号： A01K67/0278 ,  A01K67/0271 ,  A01K67/0275 ,  A01K2207/12 ,  A01K2207/15 ,  A01K2217/052 ,  A01K2217/072 ,  A01K2227/105 ,  A01K2267/0375 ,  A01K2267/0393 ,  A61D19/04 ,  C12N9/12 ,  C12N9/1205 ,  C12N15/8509 ,  C12N15/86 ,  C12N15/873 ,  C12N2015/8527 ,  C12N2740/15043 ,  C12N2740/16043 ,  C12N2799/027

摘要： A lentiviral vector was used to produce non-human animals that express human sFLT1 specifically in the murine placenta, to provide model animals of diseases such as pregnancy-induced hypertension syndrome that are close to the clinical conditions, methods for producing the model animals, methods of screening for candidate compounds as therapeutic agents for diseases such as pregnancy-induced hypertension syndrome by using the model animals, and therapeutic agents for diseases such as pregnancy-induced hypertension syndrome. As a result, the model animals were found to exhibit symptoms that are very close to the clinical conditions in human, which are presentation of hypertension as well as placental insufficiency, intrauterine growth retardation, glomerulosclerosis, and proteinuria during pregnancy, and improvement of those symptoms postpartum. Furthermore, when pravastatin was administered to this model animal, it was found that diseases such as pregnancy-induced hypertension syndrome were improved by the activation of placenta-derived growth factor (PIGF) which antagonizes sFLT1.

公开(公告)号：US20130117868A1

公开(公告)日：2013-05-09

申请号：US13516317

申请日：2010-12-03

申请人： James Cao ,  Karen Chandross ,  Kyriakos D. Economides ,  Harry Gregory Polites ,  Daniel Weinstock ,  Xiaoyou Ying

发明人： James Cao ,  Karen Chandross ,  Kyriakos D. Economides ,  Harry Gregory Polites ,  Daniel Weinstock ,  Xiaoyou Ying

IPC分类号： A01K67/027

CPC分类号： A01K67/0275 ,  A01K2217/052 ,  A01K2217/206 ,  A01K2227/105 ,  A01K2267/03 ,  A01K2267/0393 ,  C12N15/8509

摘要： A Myelin Basic Protein-luciferase bioimaging noninvasive model to visualize and quantify demyelination and remyelination events in the CNS at transcriptional level in vivo is provided. Luciferase-expressing transgenic animals were generated with myelin basic protein (MBP) promoter coupled to firefly luciferase reporter. The MBP-luci bioimaging model provides a means to monitor myelination status and the efficacy of a remyelination modulating test compound. An advantage of bioimaging is that a subject in a longitudinal study can serve as its own control. The same subject can be tracked over a demyelination and remyelination process continuously over a period of at least 10 weeks. This model enables normalization of individual animal imaging response and provides quality data with considerably reduced variance. In addition, because cohorts of animals need not be sacrificed at different time points, reduction in the number necessary for a compound efficacy study is possible.

摘要翻译： 提供了髓鞘碱性蛋白 - 萤光素酶生物成像非侵入性模型，以在转录水平的体内可视化和定量CNS中的脱髓鞘和髓鞘再生事件。 荧光素酶表达的转基因动物用与萤火虫荧光素酶报告基因偶联的髓鞘碱性蛋白（MBP）启动子产生。 MBP-luci生物成像模型提供了一种监测髓鞘形态和再髓核调节化合物的功效的方法。 生物成像的一个优点是，纵向研究中的主题可以作为自己的控制。 可以在至少10周的时间内持续跟踪脱髓鞘和髓鞘再生过程。 该模型可以实现个体动物成像响应的归一化，并提供具有显着降低的差异的质量数据。 另外，由于不需要在不同时间点牺牲动物群，所以可以减少化合物功效研究所需的数量。

公开(公告)号：US20130024955A1

公开(公告)日：2013-01-24

申请号：US13578183

申请日：2011-03-31

申请人： Hyon El Choy ,  Seok-Yong Choi

发明人： Hyon El Choy ,  Seok-Yong Choi

IPC分类号： A61K49/00 ,  C12N15/12

CPC分类号： G01N33/5014 ,  A01K67/0275 ,  A01K2217/052 ,  A01K2227/40 ,  A01K2267/0393 ,  G01N33/5088

摘要： The present invention relates to a method for determining a genotoxicity of a test substance, comprising the steps of: (a) transforming a fish with a nucleotide sequence encoding a non-fluorescent fluorescence protein with a mutation; (b) treating a test substance to the transformed fish; and (c) measuring a fluorescence in the test substance-treated fish, wherein the fluorescence is generated by reversion of the non-fluorescent fluorescence protein to the fluorescence protein due to a back mutation of the nucleotide sequence in the test substance-treated fish. According to the present invention, MutaFish system, Zebrafish (Brachydanio rerio) line, in which the fluorescence protein variant, preferably wild type EGFP variant (EGFPmut) is transformed, provides a much excellent animal system for measuring a genotoxicity of a test substance via production of a fluorescent fry larvae generated through reversion of the fluorescent protein variant caused by treatment of the test substance. Accordingly, the MutaFish system of the present invention may determine a genotoxicity of a test substance in much more simple and high-throughput manner in an eukaryote to which the conventional methods (e.g., Ames test) may be not applied.

摘要翻译： 本发明涉及测定物质的基因毒性的方法，其特征在于，包括以下步骤：（a）用编码具有突变的非荧光荧光蛋白的核苷酸序列转化鱼; （b）将经检测的物质处理至经转化的鱼; 和（c）测定被检物质处理过的鱼中的荧光，其中，由于检测物质处理的鱼中的核苷酸序列的反向突变，通过将非荧光荧光蛋白逆转为荧光蛋白而产生荧光。 根据本发明，其中荧光蛋白变体，优选野生型EGFP变体（EGFPmut）被转化的MutaFish系统斑马鱼（Brachydanio rerio）系提供了非常优异的用于通过生产测量测试物质的基因毒性的动物系统 通过由测试物质的处理引起的荧光蛋白质变体的逆转产生的荧光鱼苗。 因此，本发明的MutaFish系统可以以更加简单和高通量的方式在常规方法（例如Ames测试）中不能应用的真核细胞中确定测试物质的基因毒性。

公开(公告)号：US20130019325A1

公开(公告)日：2013-01-17

申请号：US13577565

申请日：2011-03-17

申请人： Karl Deisseroth ,  Feng Zhang ,  Viviana Gradinaru

发明人： Karl Deisseroth ,  Feng Zhang ,  Viviana Gradinaru

IPC分类号： C12N5/10 ,  A01K67/00 ,  C12N13/00

CPC分类号： C07K14/405 ,  A01K67/0271 ,  A01K67/0275 ,  A01K2207/05 ,  A01K2227/105 ,  A01K2267/0393 ,  A61K38/00 ,  A61K48/00 ,  A61N5/0601 ,  A61N5/062 ,  A61N2005/0626 ,  A61N2005/063 ,  A61N2005/0651 ,  A61N2005/0663 ,  C07K7/06 ,  C07K7/08 ,  C07K14/00 ,  C07K19/00 ,  C07K2319/01 ,  C12N15/86 ,  C12N2750/14143 ,  C12N2799/025

摘要： Disclosed are polynucleotides and methods for expressing light activated proteins in animal cells and altering an action potential of the cells by optical stimulation. The disclosure also provides animal cells and non-human animals comprising cells expressing the light-activated proteins.

摘要翻译： 公开了用于在动物细胞中表达光活化蛋白的多核苷酸和方法，并通过光学刺激改变细胞的动作电位。 本公开还提供包含表达光活化蛋白的细胞的动物细胞和非人动物。

公开(公告)号：US20120272349A1

公开(公告)日：2012-10-25

申请号：US13513100

申请日：2010-11-30

申请人： Takahiro Ochiya ,  Masaki Kawamata

发明人： Takahiro Ochiya ,  Masaki Kawamata

IPC分类号： C12N15/85 ,  A01K67/027 ,  C12N5/02

CPC分类号： C12N15/873 ,  A01K67/0271 ,  A01K67/0276 ,  A01K2217/00 ,  A01K2217/052 ,  A01K2227/105 ,  A01K2267/0393 ,  C07K2319/81 ,  C12N5/0606 ,  C12N9/22 ,  C12N15/85 ,  C12N15/8509 ,  C12N2500/30 ,  C12N2501/065 ,  C12N2501/15 ,  C12N2501/998 ,  C12N2501/999

摘要： The present invention provides a preparation method of a chimeric embryo and a chimeric rat, which is characterized by contacting a rat pluripotent stem cell and a host embryo in the presence of an ES cell differentiation inhibitor. The method includes (a) a step for contacting a fertilized host embryo collected from a female rat and a rat pluripotent stem cell in the presence of an ES cell differentiation suppressant, and (b) a step for culturing the host embryo in contact with the rat pluripotent stem cell to form a chimeric embryo.

摘要翻译： 本发明提供嵌合胚胎和嵌合大鼠的制备方法，其特征在于在ES细胞分化抑制剂存在下使大鼠多能干细胞与宿主胚胎接触。 该方法包括：（a）在ES细胞分化抑制剂存在下，将从雌性大鼠和大鼠多能干细胞收集的受精宿主胚胎接触的步骤，和（b）与所述宿主胚胎接触的步骤 大鼠多能干细胞形成嵌合胚胎。

公开(公告)号：US20120270259A1

公开(公告)日：2012-10-25

申请号：US13533117

申请日：2012-06-26

申请人： Sergey PARINOV ,  Alexander EMELYANOV

发明人： Sergey PARINOV ,  Alexander EMELYANOV

IPC分类号： C12Q1/02 ,  C12N5/071

CPC分类号： C07K14/82 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2227/40 ,  A01K2267/0331 ,  A01K2267/0393 ,  C07K5/101 ,  C07K14/461 ,  C07K2319/60 ,  C12N15/8509 ,  C12N2830/008 ,  G01N33/5011

摘要： The present invention is directed to fish whose genome has integrated therein an oncogenic nucleic acid operably linked to a promoter. Methods of making the fish and methods for their use are also provided. The fish may advantageously be utilized in methods of screening for drugs or agents that modulate oncogene-mediated neoplastic or hyperplasic transformation, or that modulate sensitivity to chemotherapy or radiation therapy Immortal tumor cells lines, methods of making immortal tumor cells lines and methods of their use are also provided.

摘要翻译： 本发明涉及其基因组整合有可操作地连接到启动子的致癌核酸的鱼。 还提供了制作鱼的方法及其使用方法。 鱼可以有利地用于筛选调节癌基因介导的肿瘤或超基因转化或调节对化学疗法或放射治疗的敏感性的药物或试剂的方法。永生肿瘤细胞系，制备不死的肿瘤细胞系的方法及其使用方法 也提供。

公开(公告)号：US20120238726A1

公开(公告)日：2012-09-20

申请号：US13484893

申请日：2012-05-31

申请人： MING-CHUNG CHANG

发明人： MING-CHUNG CHANG

IPC分类号： C07K14/28

CPC分类号： C07K14/28 ,  A01K2217/052 ,  A01K2227/706 ,  A01K2267/0393 ,  C12N15/8509

摘要： The present invention discloses a mutant blue fluorescent protein (BFP), mutated by an error-prone PCR method or a DNA shuffling method with using a BFPvv D7 of SEQ ID NO:2 as parents, obtained from a wild type blue fluorescent protein BfgV of SEQ ID NO:1, obtained from Vibrio vulnificus, wherein a set of mutation positions of the mutant BFP corresponding to SEQ ID NO:2 comprises position 176 and position 178. In a preferred embodiment, the set of mutation positions of the mutant BFP corresponding to SEQ ID NO:2 comprises a S176R mutation or a V178I mutation. Moreover, methods of using the blue fluorescent proteins from Vibrio vulnificus for fluorescence resonance energy transfer (FRET) and a blue fluorescent fish are also provided.

摘要翻译： 本发明公开了一种突变型蓝色荧光蛋白（BFP），其通过易错PCR法或使用SEQ ID NO：2的BFPvv D7作为亲本的DNA改组法从野生型蓝色荧光蛋白BfgV获得突变 从创伤弧菌获得的SEQ ID NO：1，其中与SEQ ID NO：2相对应的突变体BFP的一组突变位置包含位置176和位置178.在优选实施方案中，突变体BFP的突变位置集合对应 至SEQ ID NO：2包含S176R突变或V178I突变。 此外，还提供了使用来自创伤弧菌的蓝色荧光蛋白用于荧光共振能量转移（FRET）和蓝色荧光鱼的方法。