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公开(公告)号：US5173414A
公开(公告)日：1992-12-22
申请号：US605775
申请日：1990-10-30
IPC分类号： C12N5/10 , C07K14/015 , C07K14/05 , C12N7/00 , C12N7/04 , C12N15/09 , C12N15/38 , C12N15/864 , C12N15/869 , C12R1/92
CPC分类号： C12N15/86 , C07K14/005 , C12N7/00 , C12N2710/16222 , C12N2710/16244 , C12N2710/16262 , C12N2750/14122 , C12N2750/14143
摘要： A simplified method to produce recombinant adeno-associated virus (AAV) vectors is described. The procedure involves the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr Virus (oriP), and a recombinant AAV genome. The chimeric plasmids themselves are also a part of the present invention. These EBV/AAV plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of recombinant AAV virus upon addition of wild-type, adeno-associated virus helper functions. Recombinant AAV vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit all of the attributes of viral elements produced by conventional methods.
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公开(公告)号：US5681731A
公开(公告)日：1997-10-28
申请号：US459049
申请日：1995-06-02
IPC分类号： C12N5/10 , C07K14/015 , C07K14/05 , C12N7/00 , C12N7/04 , C12N15/09 , C12N15/38 , C12N15/864 , C12N15/869 , C12R1/92 , C12N15/63 , C12N5/06 , C12N5/08 , C12N15/70
CPC分类号： C12N15/86 , C07K14/005 , C12N7/00 , C12N2710/16222 , C12N2710/16244 , C12N2710/16262 , C12N2750/14122 , C12N2750/14143
摘要： Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of viral elements produced by conventional methods.
摘要翻译： 描述了产生重组腺相关病毒（rAAV）载体的简化方法。 该方法涉及使用掺入爱泼斯坦巴尔核抗原（EBNA）基因，爱泼斯坦巴尔病毒（oriP）的潜在复制起点）和rAAV基因组的嵌合质粒。 嵌合质粒本身也是本发明的一部分。 这些质粒作为细胞中的多拷贝染色体外元件维持，如人293细胞。 携带这些EBV / AAV质粒的永久性细胞系在添加野生型腺相关病毒辅助功能后被诱导产生大量的rAAV。 以这种方式产生的载体能够将外源基因转导到其他人细胞系中，并显示通过常规方法产生的病毒元件的属性。
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公开(公告)号：US5780280A
公开(公告)日：1998-07-14
申请号：US459352
申请日：1995-06-02
IPC分类号： C12N5/10 , C07K14/015 , C07K14/05 , C12N7/00 , C12N7/04 , C12N15/09 , C12N15/38 , C12N15/864 , C12N15/869 , C12R1/92 , C12N15/64 , C12N15/86
CPC分类号： C12N15/86 , C07K14/005 , C12N7/00 , C12N2710/16222 , C12N2710/16244 , C12N2710/16262 , C12N2750/14122 , C12N2750/14143
摘要： Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of viral elements produced by conventional methods.
摘要翻译： 描述了产生重组腺相关病毒（rAAV）载体的简化方法。 该方法涉及使用掺入爱泼斯坦巴尔核抗原（EBNA）基因，爱泼斯坦巴尔病毒（oriP）的潜在复制起点）和rAAV基因组的嵌合质粒。 嵌合质粒本身也是本发明的一部分。 这些质粒作为细胞中的多拷贝染色体外元件维持，如人293细胞。 携带这些EBV / AAV质粒的永久性细胞系在添加野生型腺相关病毒辅助功能后被诱导产生大量的rAAV。 以这种方式产生的载体能够将外源基因转导到其他人细胞系中，并显示通过常规方法产生的病毒元件的属性。
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4.发明授权
公开(公告)号：US5691176A
公开(公告)日：1997-11-25
申请号：US459091
申请日：1995-06-02
IPC分类号： C12N5/10 , C07K14/015 , C07K14/05 , C12N7/00 , C12N7/04 , C12N15/09 , C12N15/38 , C12N15/864 , C12N15/869 , C12R1/92 , C12N15/00 , C12N5/06 , C12N7/01 , C12N15/35
CPC分类号： C12N15/86 , C07K14/005 , C12N7/00 , C12N2710/16222 , C12N2710/16244 , C12N2710/16262 , C12N2750/14122 , C12N2750/14143
摘要： Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of viral elements produced by conventional methods.
摘要翻译： 描述了产生重组腺相关病毒（rAAV）载体的简化方法。 该方法涉及使用掺入爱泼斯坦巴尔核抗原（EBNA）基因，爱泼斯坦巴尔病毒（oriP）的潜在复制起点）和rAAV基因组的嵌合质粒。 嵌合质粒自身也是本发明的一部分。 这些质粒作为细胞中的多拷贝染色体外元件维持，如人293细胞。 携带这些EBV / AAV质粒的永久性细胞系在添加野生型腺相关病毒辅助功能后被诱导产生大量的rAAV。 以这种方式产生的载体能够将外源基因转导到其他人细胞系中，并显示通过常规方法产生的病毒元件的属性。
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公开(公告)号：US5589377A
公开(公告)日：1996-12-31
申请号：US490095
申请日：1995-06-06
IPC分类号： C12N5/10 , C07K14/015 , C07K14/05 , C12N7/00 , C12N7/04 , C12N15/09 , C12N15/38 , C12N15/864 , C12N15/869 , C12R1/92 , C12N7/01 , C12N15/86
CPC分类号： C12N15/86 , C07K14/005 , C12N7/00 , C12N2710/16222 , C12N2710/16244 , C12N2710/16262 , C12N2750/14122 , C12N2750/14143
摘要： Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The AAV rep and cap genes are combined in a single recombinant adenovirus vector, i.e., a rep/cap adenovirus vector which combines in a single vector all complementing functions required for rAAV production. The rep/cap vectors of the present invention comprise: genes encoding helper functions, preferably from adenovirus, sufficient to permit AAV replication; and AAV cap and rep genes. In adenovirus rep/cap vectors the rep and cap genes may, for example, replace the adenovirus E3 gene or the adenovirus E1a and E1b genes.
摘要翻译： 描述了产生重组腺相关病毒（rAAV）载体的简化方法。 AAV rep和cap基因在单个重组腺病毒载体中组合，即rep / cap腺病毒载体，其以单个载体结合rAAV产生所需的所有补体功能。 本发明的rep / cap载体包括：编码辅助功能的基因，优选来自腺病毒，足以允许AAV复制; 和AAV cap和rep基因。 在腺病毒rep / cap载体中，rep和cap基因可以例如替代腺病毒E3基因或腺病毒E1a和E1b基因。
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公开(公告)号：US5354678A
公开(公告)日：1994-10-11
申请号：US993776
申请日：1992-12-21
IPC分类号： C12N5/10 , C07K14/015 , C07K14/05 , C12N7/00 , C12N7/04 , C12N15/09 , C12N15/38 , C12N15/864 , C12N15/869 , C12R1/92 , C12N7/01 , C12N15/86
CPC分类号： C12N15/86 , C07K14/005 , C12N7/00 , C12N2710/16222 , C12N2710/16244 , C12N2710/16262 , C12N2750/14122 , C12N2750/14143
摘要： Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of vital elements produced by conventional methods.
摘要翻译： 描述了产生重组腺相关病毒（rAAV）载体的简化方法。 该方法涉及使用掺入爱泼斯坦巴尔核抗原（EBNA）基因的嵌合质粒，爱泼斯坦巴尔病毒（oriP）的潜在复制起点和rAAV基因组。 嵌合质粒本身也是本发明的一部分。 这些质粒作为细胞中的多拷贝染色体外元件维持，如人293细胞。 携带这些EBV / AAV质粒的永久性细胞系在添加野生型腺相关病毒辅助功能后被诱导产生大量的rAAV。 以这种方式产生的载体能够将外源基因转导到其他人细胞系中，并显示通过常规方法产生的重要元件的属性。
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7.发明申请DIFFERENTIATION OF PRIMATE PLURIPOTENT STEM CELLS TO HEMATOPOIETIC LINEAGE CELLS 审中-公开将原始细胞干细胞分化为HEMATOPOIETIC线细胞公开(公告)号：US20120107931A1
公开(公告)日：2012-05-03
申请号：US13312349
申请日：2011-12-06
IPC分类号： C12N5/071
CPC分类号： C12N5/0639 , A61K2039/5154 , C12N2501/02 , C12N2501/125 , C12N2501/155 , C12N2501/165 , C12N2501/22 , C12N2501/23 , C12N2501/24 , C12N2501/52 , C12N2506/02
摘要： The invention provides methods of differentiating primate pluripotent stem cells into cells of hematopoeitic lineage. The invention further provides hematopoietic lineage cells differentiated from primate pluripotent stem cells, as well as methods of using the same and kits comprising the same.
摘要翻译： 本发明提供了将灵长类动物多能干细胞分化成血细胞谱系细胞的方法。 本发明还提供了从灵长类动物多能干细胞分化的造血谱系细胞，以及使用它们的方法和包含该成分的试剂盒。
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公开(公告)号：US5807686A
公开(公告)日：1998-09-15
申请号：US482318
申请日：1995-06-06
申请人： John E. Wagner , Jane S. Lebkowski
发明人： John E. Wagner , Jane S. Lebkowski
IPC分类号： A61K35/14 , A61K35/28 , C12N5/0789 , G01N33/53 , G01N33/567 , C12N5/00 , C12N15/00
CPC分类号： C12N5/0647 , A61K35/28 , C12N2501/39 , C12N2502/1394
摘要： A cell population which is composed of cells bearing the stem cell marker CD34 and which are small in size and have little granulation are obtained by separating low density mononuclear hematopoietic cells according to size and then selecting for CD34.sup.+ cells in the smallest size fraction. The size of the cell population corresponds to that obtained at a flow rate of 25-29 ml/min in a counterflow elutriation method using a rotor equivalent to Beckman JE 5.0 spun at 900.times.g. This population of cells consists essentially of very early progenitor cells and the cells are useful in autologous bone marrow transplantation as well as gene therapy.
摘要翻译： 通过根据大小分离低密度单核造血细胞，然后选择最小尺寸级分的CD34 +细胞，获得由携带干细胞标志物CD34的细胞组成并且尺寸小且肉芽少的细胞群。 细胞群体的大小对应于使用等效于以900xg旋转的Beckman JE 5.0的转子的逆流淘析方法，以25-29ml / min的流速获得的大小。 这种细胞群基本上由非常早期的祖细胞组成，并且该细胞可用于自体骨髓移植以及基因治疗。
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9.发明授权Adeno-associated viral liposomes and their use in transfecting dendritic cells to stimulate specific immunity 失效腺相关病毒脂质体及其在转染树突细胞中的用途，以刺激特异性免疫公开(公告)号：US06652850B1
公开(公告)日：2003-11-25
申请号：US09000003
申请日：1998-06-15
申请人： Ramila Philip , Jane S. Lebkowski
发明人： Ramila Philip , Jane S. Lebkowski
IPC分类号： A61K4800
CPC分类号： C12N15/86 , A61K9/1272 , A61K47/6901 , A61K47/6911 , A61K48/00 , C07K14/47 , C07K14/55 , C07K14/70503 , C12N15/88 , C12N2710/10343 , C12N2750/14143
摘要： A composition for genetic manipulation of cells comprises a liposome comprised of lipid material, and adeno-associated viral (AAV) material. Typically, the AAV material is plasmid, and comprises one or more terminal repeats of the AAV genome. Methods are disclosed for introducing DNA into cells using AAV/liposome complexes. The DNA is introduced and expressed in'stem cells, T cells, primary tumor cells, tumor cell lines and dendritic cells or other antigen-presenting cells. Such transfected cells are used in therapeutic methods to treat subjects with cancer or microbial infections. Dendritic cells with DNA encoding tumor or viral antigens and are used to treat subjects with tumors or viral infections by administration in vivo or by activation of antigen-specific lymphocytes ex vivo followed by administration of those lymphocytes to the subject.
摘要翻译： 用于细胞遗传操作的组合物包括由脂质材料和腺相关病毒（AAV）材料组成的脂质体。 通常，AAV材料是质粒，并且包含AAV基因组的一个或多个末端重复序列。 公开了使用AAV /脂质体复合物将DNA引入细胞的方法。 引入DNA并在干细胞，T细胞，原代肿瘤细胞，肿瘤细胞系和树突状细胞或其他抗原呈递细胞中表达。 这种转染的细胞用于治疗患有癌症或微生物感染的受试者的治疗方法。 具有编码肿瘤或病毒抗原的DNA的树突状细胞用于通过在体内施用或通过在体外激活抗原特异性淋巴细胞来治疗患有肿瘤或病毒感染的受试者，然后向受试者施用那些淋巴细胞。
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公开(公告)号：US06576464B2
公开(公告)日：2003-06-10
申请号：US09783203
申请日：2001-02-13
申请人： Joseph D. Gold , Jane S. Lebkowski
发明人： Joseph D. Gold , Jane S. Lebkowski
IPC分类号： C12N500
CPC分类号： C12N15/85 , A61K35/12 , C12N5/0606 , C12N2502/13 , C12N2510/00
摘要： This invention provides a system for producing differentiated cells from a stem cell population for use wherever a relatively homogenous cell population is desirable. The cells contain an effector gene under control of a transcriptional control element (such as the TERT promoter) that causes the gene to be expressed in relatively undifferentiated cells in the population. Expression of the effector gene results in depletion of undifferentiated cells, or expression of a marker that can be used to remove them later. Suitable effector sequences encode a toxin, a protein that induces apoptosis, a cell-surface antigen, or an enzyme (such as thymidine kinase) that converts a prodrug into a substance that is lethal to the cell. The differentiated cell populations produced according to this disclosure are suitable for use in tissue regeneration, and non-therapeutic applications such as drug screening.
摘要翻译： 本发明提供了一种用于从干细胞群体产生分化细胞的系统，用于需要相对均匀细胞群体的地方。 细胞含有在转录控制元件（例如TERT启动子）的控制下的效应基因，其导致基因在群体中相对未分化的细胞中表达。 效应基因的表达导致未分化细胞的消耗或可以用于稍后去除它们的标记物的表达。 合适的效应序列编码毒素，诱导凋亡的蛋白质，细胞表面抗原或将前药转化为对细胞致死的物质的酶（例如胸苷激酶）。 根据本公开生产的分化细胞群体适用于组织再生和非治疗应用，例如药物筛选。
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