公开(公告)号：US08338168B2

公开(公告)日：2012-12-25

申请号：US10512474

申请日：2003-04-25

申请人： Didier Trono ,  Francois-Loic Cosset ,  Virginie Sandrin ,  Bertrand Boson ,  Didier Negre ,  Patrick Salmon

发明人： Didier Trono ,  Francois-Loic Cosset ,  Virginie Sandrin ,  Bertrand Boson ,  Didier Negre ,  Patrick Salmon

IPC分类号： C12N15/00 ,  C12N15/64

CPC分类号： C12N15/86 ,  A61K48/00 ,  C07K14/005 ,  C07K2319/00 ,  C07K2319/03 ,  C12N7/045 ,  C12N2510/02 ,  C12N2740/13022 ,  C12N2740/13043 ,  C12N2740/15043 ,  C12N2740/15045 ,  C12N2810/60 ,  C12N2830/50

摘要： The present invention provides improved chimeric glycoproteins (GPs) and improved lentiviral vectors pseudotyped with those glycoproteins. Also provided are methods and compositions for making such glycoproteins and vectors, and improved methods of in vitro and in vivo transduction of cells with such vectors. Improved chimeric GPs encode the extracellular and transmembrane domains of GALV or RD114 GPs fused to the cytoplasmic tail of MLV-A GP. Vectors pseudotyped with these GAL V/TR and RD 114/TR GP chimeras have significantly higher titers than vectors coated with the parental GPs. Additionally, RD114/TR-pseudotyped vectors are efficiently concentrated and are resistant to inactivation induced by the complement of both human and macaque sera. RD114 GP-pseudotyped lentiviral vectors have particular utility for in vivo gene transfer applications.

摘要翻译： 本发明提供改进的嵌合糖蛋白（GP）和用这些糖蛋白假型分类的改良的慢病毒载体。 还提供了用于制备这种糖蛋白和载体的方法和组合物，以及用这些载体体外和体内转导细胞的改进方法。 改进的嵌合GP编码与MLV-A GP的细胞质尾融合的GALV或RD114GP的细胞外和跨膜结构域。 与这些GAL V / TR和RD 114 / TR GP嵌合体假型分离的载体具有比用亲本GP包被的载体更高的滴度。 此外，RD114 / TR-假型载体有效浓缩，并且对由人和猕猴血清的补体诱导的失活具有抗性。 RD114 GP-假型慢病毒载体对于体内基因转移应用具有特殊的用途。

公开(公告)号：US20100062524A1

公开(公告)日：2010-03-11

申请号：US12578346

申请日：2009-10-13

申请人： Didier Trono ,  Romain N. Zufferey

发明人： Didier Trono ,  Romain N. Zufferey

IPC分类号： C12N15/63

CPC分类号： C12N15/86 ,  A61K48/00 ,  C12N7/00 ,  C12N2740/15043 ,  C12N2740/16043 ,  C12N2740/16052 ,  C12N2740/16061 ,  C12N2830/38 ,  C12N2830/48 ,  C12N2830/50 ,  C12N2840/203 ,  C12N2840/44

摘要： The present invention provides HIV-derived lentivectors which are multiply modified to create highly safe, efficient, and potent vectors for expressing transgenes for gene therapy. The lentiviral vectors comprise various combinations of an inactive central polypurine tract, a stuffer sequence, which may encode drug susceptibility genes, and a mutated hairpin in the 5′ leader sequence that substantially abolishes replication. These elements are provided in conjunction with other features of lentiviral vectors, such as a self-inactivating configuration for biosaftey and promoters such as the EF1α promoter as one example. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element. These vectors therefore provide useful tools for genetic treatments for inherited and acquired disorders, gene-therapies for cancers and other disease, the creation of industrial and experimental production systems utilizing transformed cells, as well as for the study of basic cellular and genetic processes.

摘要翻译： 本发明提供了HIV衍生的慢病毒药物，其被多重修饰以产生用于表达用于基因治疗的转基因的高度安全，有效和有效的载体。 慢病毒载体包括不规则的中枢多巴胺，可编码药物敏感性基因的填充序列和基本上消除复制的5'前导序列中突变的发夹的各种组合。 这些元件与慢病毒载体的其它特征结合提供，例如用于生物卫生的自灭活构型和启动子如EF1α启动子作为一个实例。 还描述了另外的启动子。 载体还可以包含额外的转录增强元件，例如木吸虫肝炎病毒转录后调节元件。 因此，这些载体为继承和获得性疾病的遗传治疗，癌症和其他疾病的基因治疗，利用转化细胞的工业和实验生产系统的建立以及基础细胞和遗传过程的研究提供了有用的工具。

公开(公告)号：US20050014166A1

公开(公告)日：2005-01-20

申请号：US10720987

申请日：2003-11-24

申请人： Didier Trono ,  Maciej Wiznerowicz

发明人： Didier Trono ,  Maciej Wiznerowicz

IPC分类号： A61K48/00 ,  C12N15/11 ,  C12N15/113 ,  C12N15/86 ,  C12N15/867 ,  C12Q1/68

CPC分类号： C12N15/8509 ,  A01K2217/05 ,  A61D19/04 ,  C12N15/111 ,  C12N15/113 ,  C12N15/86 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2320/32 ,  C12N2320/50 ,  C12N2740/16043 ,  C12N2800/30 ,  C12N2830/003 ,  C12N2830/005 ,  C12N2830/008 ,  C12N2830/48 ,  C12N2830/50 ,  C12N2840/203

摘要： The present invention provides compositions and methods of modulating or regulating eukaryotic gene expression through the controlled or regulated expression of polynucleotide constructs that encode siRNA or other desired exogenous nucleic acids or proteins. Such constructs, and additional elements of the system may be transfected into the cells of interest and the expression of the siRNA, and hence the expression of the target gene of the siRNA, may be controlled through the administration of a compound to the cell, such as a small molecule or drug. Lentivirus vectors are employed in some embodiments of the invention including the generation of conditional knockdown animals.

摘要翻译： 本发明提供了通过控制或调控表达编码siRNA或其它所需外源核酸或蛋白质的多核苷酸构建体来调节或调节真核基因表达的组合物和方法。 可以将这样的构建体和系统的另外的元件转染到感兴趣的细胞中，并且可以通过向细胞施用化合物来控制siRNA的表达，从而siRNA的靶基因的表达，例如 作为小分子或药物。 在本发明的一些实施方案中使用慢病毒载体，包括产生条件性敲除动物。

公开(公告)号：US5358856A

公开(公告)日：1994-10-25

申请号：US887223

申请日：1992-05-18

申请人： David Baltimore ,  Didier Trono

发明人： David Baltimore ,  Didier Trono

IPC分类号： C12N15/85 ,  C12N5/10 ,  C12P21/02

CPC分类号： C12N15/85 ,  C12N2840/203

摘要： A construct which includes a cap independent 5' noncoding region of viral or cellular origin and a nucleotide sequence of interest, which is located downstream of the noncoding region. A method of producing a protein or a polypeptide of interest by introducing the construct, including a nucleotide sequence encoding the protein or the polypeptide of interest, into mammalian cells is also described. In one embodiment, the construct comprises all or a portion of the poliovirus cap independent 5' noncoding region and a nucleotide sequence encoding a protein or a polypeptide of interest.

摘要翻译： 包括病毒或细胞来源的帽独立的5'非编码区和感兴趣的核苷酸序列的构建体，其位于非编码区的下游。 还描述了通过将编码所述蛋白质或目标多肽的核苷酸序列引入哺乳动物细胞的构建体来产生目的蛋白质或多肽的方法。 在一个实施方案中，构建体包含脊髓灰质炎病毒帽独立的5'非编码区的全部或一部分和编码感兴趣的蛋白质或多肽的核苷酸序列。

公开(公告)号：US08748169B2

公开(公告)日：2014-06-10

申请号：US11680414

申请日：2007-02-28

申请人： Didier Trono ,  Maciej Wiznerowicz

发明人： Didier Trono ,  Maciej Wiznerowicz

IPC分类号： C12N15/63 ,  C12N15/85

CPC分类号： C12N15/86 ,  A61K48/00 ,  C07K14/55 ,  C07K14/70503 ,  C07K16/00 ,  C07K2317/622 ,  C07K2319/30 ,  C12N2740/15043 ,  C12N2740/16043 ,  C12N2800/30 ,  C12N2830/00 ,  C12N2830/002 ,  C12N2830/008 ,  C12N2830/30 ,  C12N2830/48 ,  C12N2830/50 ,  C12N2830/85 ,  C12N2840/20 ,  C12N2840/44

摘要： The present invention provides HIV-derived lentivectors which are safe, highly efficient, and very potent for expressing transgenes for human gene therapy, especially, in human hematopoietic progenitor cells as well as in all other blood cell derivatives. The lentiviral vectors comprise promoters active to promote expression specific to cell types or tissues. Further, promoters are providing that are amenable to control by activators, enhancers, or repressors. These vectors are in a self-inactivating configuration for biosaftey. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element, without any decrease in the specificity or control exerted by the promoters. These vectors therefore provide useful tools for genetic treatments such as inherited and acquired lympho-hematological disorders, gene-therapies for cancers especially the hematological cancers, as well as for the study of hematopoiesis via lentivector-mediated modification of human HSCs.

摘要翻译： 本发明提供了用于人基因治疗特别是人类造血祖细胞以及所有其他血液细胞衍生物中表达转基因的安全，高效和非常有效的HIV衍生的慢病毒药物。 慢病毒载体包含促进对细胞类型或组织特异性表达的活性的启动子。 此外，提供可由活化剂，增强剂或阻遏物控制的启动子。 这些载体是生物燃料的自灭活配置。 还描述了另外的启动子。 载体还可以包含额外的转录增强元件，例如木卡因肝炎病毒转录后调节元件，而启动子所引起的特异性或对照的任何降低。 因此，这些载体为遗传治疗提供了有用的工具，例如遗传和获得性淋巴 - 血液学障碍，癌症特别是血液癌症的基因治疗，以及通过慢性介导的人类HSC修饰研究造血功能。

公开(公告)号：US06284469B1

公开(公告)日：2001-09-04

申请号：US09597606

申请日：2000-06-20

申请人： Thomas J. Hope ,  Romain Zufferey ,  Didier Trono ,  John Edward Donello

发明人： Thomas J. Hope ,  Romain Zufferey ,  Didier Trono ,  John Edward Donello

IPC分类号： C12Q168

CPC分类号： C07K14/005 ,  C12N15/8216 ,  C12N2730/10122 ,  C12Q1/68 ,  C12Q1/6897

摘要： A cis-acting posttranscriptional regulatory element (PRE) useful for efficient RNA export of RNA is provided. The element, termed WPRE, is originally derived from woodchuck hepatitis virus. The invention also provides a method for enhancing the expression of transgenes by insertion of the WPRE nucleic acid sequences in operably linkage with the transgene.

摘要翻译： 提供了一种用于有效RNA RNA输出的顺式作用后转录调控元件（PRE）。 称为WPRE的元件最初来源于土拨鼠肝炎病毒。 本发明还提供了通过插入与转基因可操作地连接的WPRE核酸序列来增强转基因表达的方法。

公开(公告)号：US08329462B2

公开(公告)日：2012-12-11

申请号：US12537789

申请日：2009-08-07

申请人： Didier Trono ,  Patrick Salmon

发明人： Didier Trono ,  Patrick Salmon

IPC分类号： C12N15/867 ,  C12N5/07 ,  C12N5/071 ,  C12N5/10

CPC分类号： A61K48/0058 ,  A61K31/7105 ,  A61K35/17 ,  A61K48/00 ,  C07K14/5434 ,  C07K14/55 ,  C07K14/70503 ,  C07K14/7051 ,  C07K14/755 ,  C07K16/00 ,  C07K16/46 ,  C07K2317/622 ,  C12N7/00 ,  C12N15/86 ,  C12N2740/15043 ,  C12N2740/15045 ,  C12N2740/16043 ,  C12N2740/16045 ,  C12N2740/16211 ,  C12N2800/108 ,  C12N2830/50 ,  C12N2840/20

摘要： The present invention provides HIV-derived lentivectors which are safe, highly efficient, and very potent for expressing transgenes for human gene therapy, especially, in human hematopoietic progenitor cells as well as in all other blood cell derivatives. The lentiviral vectors comprise a self-inactivating configuration for biosaftey and promoters such as the EF1α promoter as one example. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element. These vectors therefore provide useful tools for genetic treatments such as inherited and acquired lympho-hematological disorders, gene-therapies for cancers especially the hematological cancers, as well as for the study of hematopoiesis via lentivector-mediated modification of human HSCs.

摘要翻译： 本发明提供了用于人基因治疗特别是人类造血祖细胞以及所有其他血液细胞衍生物中表达转基因的安全，高效和非常有效的HIV衍生的慢病毒药物。 慢病毒载体包括用于生物卫生的自灭活构型和启动子如EF1α启动子作为一个实例。 还描述了另外的启动子。 载体还可以包含额外的转录增强元件，例如木吸虫肝炎病毒转录后调节元件。 因此，这些载体为遗传治疗提供了有用的工具，例如遗传和获得性淋巴 - 血液学障碍，癌症特别是血液癌症的基因治疗，以及通过慢性介导的人类HSC修饰研究造血功能。

公开(公告)号：US07575924B2

公开(公告)日：2009-08-18

申请号：US10010081

申请日：2001-11-09

申请人： Didier Trono ,  Patrick Salmon

发明人： Didier Trono ,  Patrick Salmon

IPC分类号： C12N5/08 ,  C12N5/10 ,  C12N15/861

CPC分类号： A61K48/0058 ,  A61K31/7105 ,  A61K35/17 ,  A61K48/00 ,  C07K14/5434 ,  C07K14/55 ,  C07K14/70503 ,  C07K14/7051 ,  C07K14/755 ,  C07K16/00 ,  C07K16/46 ,  C07K2317/622 ,  C12N7/00 ,  C12N15/86 ,  C12N2740/15043 ,  C12N2740/15045 ,  C12N2740/16043 ,  C12N2740/16045 ,  C12N2740/16211 ,  C12N2800/108 ,  C12N2830/50 ,  C12N2840/20

摘要： The present invention provides HIV-derived lentivectors which are safe, highly efficient, and very potent for expressing transgenes for human gene therapy, especially, in human hematopoietic progenitor cells as well as in all other blood cell derivatives. The lentiviral vectors comprise a self-inactivating configuration for biosaftey and promoters such as the EF1α promoter as one example. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element. These vectors therefore provide useful tools for genetic treatments such as inherited and acquired lympho-hematological disorders, gene-therapies for cancers especially the hematological cancers, as well as for the study of hematopoiesis via lentivector-mediated modification of human HSCs.

摘要翻译： 本发明提供了用于人基因治疗特别是人类造血祖细胞以及所有其他血液细胞衍生物中表达转基因的安全，高效和非常有效的HIV衍生的慢病毒药物。 慢病毒载体包括用于生物卫生的自灭活构型和启动子如EF1α启动子作为一个实例。 还描述了另外的启动子。 载体还可以包含额外的转录增强元件，例如木吸虫肝炎病毒转录后调节元件。 因此，这些载体为遗传治疗提供了有用的工具，例如遗传和获得性淋巴 - 血液学障碍，癌症特别是血液癌症的基因治疗，以及通过慢性介导的人类HSC修饰研究造血功能。

公开(公告)号：US20070196922A1

公开(公告)日：2007-08-23

申请号：US11680414

申请日：2007-02-28

申请人： Didier Trono ,  Maciej Wiznerowicz

发明人： Didier Trono ,  Maciej Wiznerowicz

IPC分类号： C12N15/867 ,  C12N5/08

CPC分类号： C12N15/86 ,  A61K48/00 ,  C07K14/55 ,  C07K14/70503 ,  C07K16/00 ,  C07K2317/622 ,  C07K2319/30 ,  C12N2740/15043 ,  C12N2740/16043 ,  C12N2800/30 ,  C12N2830/00 ,  C12N2830/002 ,  C12N2830/008 ,  C12N2830/30 ,  C12N2830/48 ,  C12N2830/50 ,  C12N2830/85 ,  C12N2840/20 ,  C12N2840/44

摘要： The present invention provides HIV-derived lentivectors which are safe, highly efficient, and very potent for expressing transgenes for human gene therapy, especially, in human hematopoietic progenitor cells as well as in all other blood cell derivatives. The lentiviral vectors comprise promoters active to promote expression specific to cell types or tissues. Further, promoters are providing that are amenable to control by activators, enhancers, or repressors. These vectors are in a self-inactivating configuration for biosaftey. Additional promoters are also described. The vectors can also comprise additional transcription enhancing elements such as the wood chuck hepatitis virus post-transcriptional regulatory element, without any decrease in the specificity or control exerted by the promoters. These vectors therefore provide useful tools for genetic treatments such as inherited and acquired lympho-hematological disorders, gene-therapies for cancers especially the hematological cancers, as well as for the study of hematopoiesis via lentivector-mediated modification of human HSCs.

摘要翻译： 本发明提供了用于人基因治疗特别是人类造血祖细胞以及所有其他血液细胞衍生物中表达转基因的安全，高效和非常有效的HIV衍生的慢病毒药物。 慢病毒载体包含促进对细胞类型或组织特异性表达的活性的启动子。 此外，提供可由活化剂，增强剂或阻遏物控制的启动子。 这些载体是生物燃料的自灭活配置。 还描述了另外的启动子。 载体还可以包含额外的转录增强元件，例如木卡因肝炎病毒转录后调节元件，而启动子所引起的特异性或对照的任何降低。 因此，这些载体为遗传治疗提供了有用的工具，例如遗传和获得性淋巴 - 血液学障碍，癌症特别是血液癌症的基因治疗，以及通过慢性介导的人类HSC修饰研究造血功能。

公开(公告)号：US20060257366A1

公开(公告)日：2006-11-16

申请号：US10512474

申请日：2003-04-25

申请人： Didier Trono ,  Francois-Loic Cosset ,  Virginie Sandrin ,  Didier Negre ,  Patrick Salmon

发明人： Didier Trono ,  Francois-Loic Cosset ,  Virginie Sandrin ,  Didier Negre ,  Patrick Salmon

IPC分类号： C12N5/08 ,  A61K48/00 ,  C12N15/86

CPC分类号： C12N15/86 ,  A61K48/00 ,  C07K14/005 ,  C07K2319/00 ,  C07K2319/03 ,  C12N7/045 ,  C12N2510/02 ,  C12N2740/13022 ,  C12N2740/13043 ,  C12N2740/15043 ,  C12N2740/15045 ,  C12N2810/60 ,  C12N2830/50

摘要： The present invention provides improved chimeric glycoproteins (GPs) and improved lentiviral vectors pseudotyped with those glycoproteins. Also provided are methods and compositions for making such glycoproteins and vectors, and improved methods of in vitro and in vivo transduction of cells with such vectors. Improved chimeric GPs encode the extracellular and transmembrane domains of GALV or RD114 GPs fused to the cytoplasmic tail of MLV-A GP. Vectors pseudotyped with these GAL V/TR and RD 114/TR GP chimeras have significantly higher titers than vectors coated with the parental GPs. Additionally, RD114/TR-pseudotyped vectors are efficiently concentrated and are resistant to inactivation induced by the complement of both human and macaque sera. RD114 GP-pseudotyped lentiviral vectors have particular utility for in vivo gene transfer applications.

摘要翻译： 本发明提供改进的嵌合糖蛋白（GP）和用这些糖蛋白假型分类的改良的慢病毒载体。 还提供了用于制备这种糖蛋白和载体的方法和组合物，以及用这些载体体外和体内转导细胞的改进方法。 改进的嵌合GP编码与MLV-A GP的细胞质尾融合的GALV或RD114GP的细胞外和跨膜结构域。 与这些GAL V / TR和RD 114 / TR GP嵌合体假型分离的载体具有比用亲本GP包被的载体更高的滴度。 此外，RD114 / TR-假型载体有效浓缩，并且对由人和猕猴血清的补体诱导的失活具有抗性。 RD114 GP-假型慢病毒载体对于体内基因转移应用具有特殊的用途。