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1. 发明授权

US06432705B1 Inducible expression system 失效
标题翻译：诱导表达系统
公开(公告)号：US06432705B1
公开(公告)日：2002-08-13
申请号：US09566660
申请日：2000-05-08
申请人： Jiing-Kuan Yee , Theodore Friedmann , Shin-Tai Chen
发明人： Jiing-Kuan Yee , Theodore Friedmann , Shin-Tai Chen
IPC分类号： C12N500
CPC分类号： C12N15/86 , C12N2740/13043 , C12N2830/002 , C12N2830/006 , C12N2840/20 , C12N2840/203
摘要： The present invention features compositions and methods for the inducible expression of a polypeptide, especially a polypeptide normally cytotoxic to the eukaryotic host cell in which it is to be expressed. A nucleotide sequence encoding a polypeptide of interest is operably linked to an inducible promoter. Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription, a transcriptional activation domain, and a second ligand-binding domain that positively regulates the transcriptional activation function of the transactivator. Transcription of the nucleotide sequence under control of the inducible promoter is activated by the multi-chimeric transactivator when both the ligand that binds the first ligand-binding domain is absent and the ligand that binds the second ligand-binding domain is present. This inducible expression system is particularly useful in the expression of the cytotoxic protein VSV G for the production of pseudotyped retroviral vectors.
摘要翻译：本发明的特征在于可诱导表达多肽的组合物和方法，特别是对待其表达的真核宿主细胞通常具有细胞毒性的多肽。编码目的多肽的核苷酸序列与诱导型启动子可操作地连接。来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节转录的第一配体结合结构域，转录激活结构域和正调节反式激活因子的转录激活功能的第二配体结合结构域组成。当两个结合第一配体结合结构域的配体和不存在结合第二配体结合结构域的配体时，多重嵌合反式激活剂可激活在诱导型启动子控制下的核苷酸序列的转录。这种诱导表达系统特别可用于细胞毒素蛋白VSV G的表达以产生假型逆转录病毒载体。

2. 发明授权

US6133027A Inducible expression system 失效
标题翻译：诱导表达系统
公开(公告)号：US6133027A
公开(公告)日：2000-10-17
申请号：US693940
申请日：1996-08-07
申请人： Jiing-Kuan Yee , Theodore Friedmann , Shin-Tai Chen
发明人： Jiing-Kuan Yee , Theodore Friedmann , Shin-Tai Chen
IPC分类号： C12N15/867 , C12N15/85 , C12N15/86
CPC分类号： C12N15/86 , C12N2740/13043 , C12N2830/002 , C12N2830/006 , C12N2840/20 , C12N2840/203
摘要： The present invention features compositions and methods for the inducible expression of a polypeptide, especially a polypeptide normally cytotoxic to the eukaryotic host cell in which it is to be expressed. A nucleotide sequence encoding a polypeptide of interest is operably linked to an inducible promoter (e.g, a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription (e.g., a prokaryotic tetracycline repressor polypeptide), a transcriptional activation domain, and a second ligand-binding domain that positively regulates the transcriptional activation function of the transactivator (e.g., a ligand-binding domain of a steroid receptor, preferably an estrogen receptor (ER)). Transcription of the nucleotide sequence under control of the inducible promoter is activated by the multi-chimeric transactivator when both the ligand that binds the first ligand-binding domain (e.g., tetracycline) is absent and the ligand that binds the second ligand-binding domain (e.g., a steroid) is present. This inducible expression system is particularly useful in the expression of the cytotoxic protein VSV G for the production of pseudotyped retroviral vectors.
摘要翻译：本发明的特征在于可诱导表达多肽的组合物和方法，特别是对待其表达的真核宿主细胞通常具有细胞毒性的多肽。编码感兴趣多肽的核苷酸序列可操作地连接到诱导型启动子（例如，由连接至多拷贝tetO的最小启动子，大肠杆菌四环素抗性操纵子Tn10的四环素阻遏物（tetR）的结合位点，）。来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节转录的第一配体结合结构域（例如原核四环素阻遏物多肽），转录激活结构域和第二配体结合结构域组成正调节反式激活因子的转录激活功能（例如，类固醇受体的配体结合结构域，优选雌激素受体（ER））。当结合第一配体结合结构域（例如四环素）的配体和结合第二配体结合结构域的配体两者都可以通过多嵌合反式激活因子激活受诱导型启动子控制下的核苷酸序列的转录（例如，类固醇）存在。这种诱导表达系统特别可用于细胞毒素蛋白VSV G的表达以产生假型逆转录病毒载体。

3. 发明授权

US5739018A Packaging cell lines for pseudotyped retroviral vectors 失效
标题翻译：包装用于假型逆转录病毒载体的细胞系
公开(公告)号：US5739018A
公开(公告)日：1998-04-14
申请号：US694652
申请日：1996-08-07
申请人： Atsushi Miyanohara , Jiing-Kuan Yee , Shin-Tai Chen , Charles Edward Prussak , Theodore Friedmann
发明人： Atsushi Miyanohara , Jiing-Kuan Yee , Shin-Tai Chen , Charles Edward Prussak , Theodore Friedmann
IPC分类号： C12N15/63 , C12N15/867 , C12N5/10 , C12N15/86
CPC分类号： C12N15/86 , C12N15/635 , C12N2740/13052 , C12N2830/002 , C12N2830/006 , C12N2830/15 , C12N2830/30 , C12N2840/20 , C12N2840/203
摘要： The present invention features packaging cell lines and recombinant retroviral particles produced thereby, particularly pseudotyped retroviral particles. Preferably, the packaging cell lines are derived from HeLa, Cf2Th, D17, MDCK, or BHK cells, most preferably from Cf2Th cells. Retroviral particles are produced by inducibly expressing an envelope protein of interest (e.g., a retroviral envelope or the envelope protein of vesicular stomatitis virus (VSV G)). Inducible expression of the envelope protein is accomplished by operably linking an envelope protein-encoding nucleotide sequence to an inducible promoter (e.g., a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli, tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription from the inducible promoter (e.g., a prokaryotic tetracycline repressor polypeptide (tetR)), a transcriptional activation domain, and a second ligand-binding domain (e.g., a ligand-binding domain of a steroid receptor, preferably an estrogen receptor (ER)).
摘要翻译：本发明的特征在于包装细胞系和由此产生的重组逆转录病毒颗粒，特别是假型逆转录病毒颗粒。优选地，包装细胞系衍生自HeLa，Cf2Th，D17，MDCK或BHK细胞，最优选来自Cf2Th细胞。逆转录病毒颗粒通过诱导表达感兴趣的包膜蛋白（例如，逆转录病毒包膜或水泡性口炎病毒的包膜蛋白（VSV G））产生。通过将包膜蛋白编码核苷酸序列可操作地连接到诱导型启动子（例如，由与tetO的多拷贝连接的最小启动子，四环素阻遏物（tetR）的结合位点）组成的启动子来实现包膜蛋白的诱导表达大肠杆菌，四环素抗性操纵子Tn10）。来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节来自诱导型启动子的转录的第一配体结合结构域（例如，原核四环素阻遏物多肽（tetR）），转录激活结构域和第二配体结合结构域（例如，类固醇受体的配体结合结构域，优选雌激素受体（ER））。

4. 发明申请

US20070212332A1 Methods for accelerating bone repair 审中-公开
标题翻译：加速骨修复的方法
公开(公告)号：US20070212332A1
公开(公告)日：2007-09-13
申请号：US11503365
申请日：2006-08-10
申请人： David Baylink , Charles Rundle , Kin-Hing Lau , Donna Strong , Shin-Tai Chen , Thomas Linkhart
发明人： David Baylink , Charles Rundle , Kin-Hing Lau , Donna Strong , Shin-Tai Chen , Thomas Linkhart
IPC分类号： A61K48/00
CPC分类号： C12N9/0083 , A61K48/005 , C07K14/51 , C12N2799/027 , C12Y114/99001
摘要： Vectors, such as retroviral vectors and transposon-based nonviral vectors, are disclosed herein that can be used to target transgene expression to the proliferating periosteal cells and cells in the marrow space after bone fracture. In one embodiment, these vectors include a human Cox-2 gene that is modified to improve mRNA stability and protein translation by truncating the 3′ untranslated region (UTR). In addition, in some embodiments, the native translation signal is replaced with an optimized Kozak sequence. These vectors can be used alone or with vectors expressing BMP2/4, FGF-2, or LMP-1 gene to repair bone fractures and increase prostaglandin secretion. Methods for identifying agents that accelerate bone repair are also disclosed.
摘要翻译：本文公开了载体，例如逆转录病毒载体和基于转座子的非病毒载体，其可以用于靶向骨骨折后骨髓空间中增殖性骨膜细胞和细胞的转基因表达。在一个实施方案中，这些载体包括通过截短3'非翻译区（UTR）修饰以改善mRNA稳定性和蛋白质翻译的人Cox-2基因。此外，在一些实施例中，用优化的Kozak序列替换天文翻译信号。这些载体可以单独使用或与使用表达BMP2 / 4，FGF-2或LMP-1基因的载体一起使用来修复骨折并增加前列腺素分泌。还公开了用于鉴定促进骨修复的药剂的方法。

5. 发明授权

US08772571B2 Compositions and methods for osteogenic gene therapy 有权
标题翻译：成骨基因治疗的组成和方法
公开(公告)号：US08772571B2
公开(公告)日：2014-07-08
申请号：US12879842
申请日：2010-09-10
申请人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
发明人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
IPC分类号： A01K67/00 , A61K48/00 , C12N5/00 , C12N15/63
CPC分类号： C07K14/50 , A01K2207/12 , A01K2267/035 , A61K38/1825 , C12N15/11 , C12N2799/027
摘要： The present disclosure provides compositions and methods for increasing bone growth and/or enhancing wound healing, for example, fracture repair. The disclosure provides recombinant nucleic acids useful for promoting bone growth. For example, the disclosure provides recombinant nucleic acids that encode a fibroblast growth factor-2 (FGF-2) analog. The disclosure also provides vectors and cells incorporating these nucleic acids, as well as FGF-2 analogs encode by them. The disclosure also provides a mouse system of bone marrow transplantation and methods for producing as well as methods for using the system. Methods for inducing division and/or inducing differentiation of a hematopoietic stem cell are also provided, as are methods for enhancing bone growth and/or wound repair (for example, fracture repair).
摘要翻译：本公开提供了用于增加骨生长和/或增强伤口愈合的组合物和方法，例如骨折修复。本公开提供了可用于促进骨生长的重组核酸。例如，本公开提供编码成纤维细胞生长因子-2（FGF-2）类似物的重组核酸。本公开还提供了掺入这些核酸的载体和细胞，以及由它们编码的FGF-2类似物。本公开还提供了骨髓移植的小鼠系统以及用于生产的方法以及使用该系统的方法。还提供诱导造血干细胞分化和/或诱导分化的方法，以及用于增强骨生长和/或伤口修复（例如，骨折修复）的方法。

6. 发明申请

US20110004947A1 COMPOSITIONS AND METHODS FOR OSTEOGENIC GENE THERAPY 有权
标题翻译：组合物和方法用于卵泡基因治疗
公开(公告)号：US20110004947A1
公开(公告)日：2011-01-06
申请号：US12879842
申请日：2010-09-10
申请人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
发明人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
IPC分类号： A01K67/00 , C12N5/071 , C12N5/0783 , C07H21/04 , C12N15/00 , G01N33/53
CPC分类号： C07K14/50 , A01K2207/12 , A01K2267/035 , A61K38/1825 , C12N15/11 , C12N2799/027
摘要： The present disclosure provides compositions and methods for increasing bone growth and/or enhancing wound healing, for example, fracture repair. The disclosure provides recombinant nucleic acids useful for promoting bone growth. For example, the disclosure provides recombinant nucleic acids that encode a fibroblast growth factor-2 (FGF-2) analog. The disclosure also provides vectors and cells incorporating these nucleic acids, as well as FGF-2 analogs encode by them. The disclosure also provides a mouse system of bone marrow transplantation and methods for producing as well as methods for using the system. Methods for inducing division and/or inducing differentiation of a hematopoietic stem cell are also provided, as are methods for enhancing bone growth and/or wound repair (for example, fracture repair).
摘要翻译：本公开提供了用于增加骨生长和/或增强伤口愈合的组合物和方法，例如骨折修复。本公开提供了可用于促进骨生长的重组核酸。例如，本公开提供编码成纤维细胞生长因子-2（FGF-2）类似物的重组核酸。本公开还提供了掺入这些核酸的载体和细胞，以及由它们编码的FGF-2类似物。本公开还提供了骨髓移植的小鼠系统以及用于生产的方法以及使用该系统的方法。还提供诱导造血干细胞分化和/或诱导分化的方法，以及用于增强骨生长和/或伤口修复（例如，骨折修复）的方法。

7. 发明授权

US07816140B2 Composition and methods for osteogenic gene therapy 有权
标题翻译：成骨基因治疗的组成和方法
公开(公告)号：US07816140B2
公开(公告)日：2010-10-19
申请号：US11452873
申请日：2006-06-13
申请人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
发明人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
IPC分类号： C12N5/00 , C12N5/02 , C12N5/10 , C12N15/63
CPC分类号： C07K14/50 , A01K2207/12 , A01K2267/035 , A61K38/1825 , C12N15/11 , C12N2799/027
摘要： The present disclosure provides compositions and methods for increasing bone growth and/or enhancing wound healing, for example, fracture repair. The disclosure provides recombinant nucleic acids useful for promoting bone growth. For example, the disclosure provides recombinant nucleic acids that encode a fibroblast growth factor-2 (FGF-2) analog. The disclosure also provides vectors and cells incorporating these nucleic acids, as well as FGF-2 analogs encode by them. The disclosure also provides a mouse system of bone marrow transplantation and methods for producing as well as methods for using the system. Methods for inducing division and/or inducing differentiation of a hematopoietic stem cell are also provided, as are methods for enhancing bone growth and/or wound repair (for example, fracture repair).
摘要翻译：本公开提供了用于增加骨生长和/或增强伤口愈合的组合物和方法，例如骨折修复。本公开提供了可用于促进骨生长的重组核酸。例如，本公开提供编码成纤维细胞生长因子-2（FGF-2）类似物的重组核酸。本公开还提供了掺入这些核酸的载体和细胞，以及由它们编码的FGF-2类似物。本公开还提供了骨髓移植的小鼠系统以及用于生产的方法以及使用该系统的方法。还提供诱导造血干细胞分化和/或诱导分化的方法，以及用于增强骨生长和/或伤口修复（例如，骨折修复）的方法。

8. 发明申请

US20100221232A1 COMPOSITION AND METHODS FOR OSTEOGENIC GENE THERAPY 有权
标题翻译：组合物和方法用于卵泡基因治疗
公开(公告)号：US20100221232A1
公开(公告)日：2010-09-02
申请号：US11452873
申请日：2006-06-13
申请人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
发明人： Kin-Hing William Lau , David J. Baylink , Susan L. Hall , Shin-Tai Chen , Subburaman Mohan
IPC分类号： A61K35/28 , A61P19/08 , A61P19/10 , A61K35/14
CPC分类号： C07K14/50 , A01K2207/12 , A01K2267/035 , A61K38/1825 , C12N15/11 , C12N2799/027
摘要： The present disclosure provides compositions and methods for increasing bone growth and/or enhancing wound healing, for example, fracture repair. The disclosure provides recombinant nucleic acids useful for promoting bone growth. For example, the disclosure provides recombinant nucleic acids that encode a fibroblast growth factor-2 (FGF-2) analog. The disclosure also provides vectors and cells incorporating these nucleic acids, as well as FGF-2 analogs encode by them. The disclosure also provides a mouse system of bone marrow transplantation and methods for producing as well as methods for using the system. Methods for inducing division and/or inducing differentiation of a hematopoietic stem cell are also provided, as are methods for enhancing bone growth and/or wound repair (for example, fracture repair).
摘要翻译：本公开提供了用于增加骨生长和/或增强伤口愈合的组合物和方法，例如骨折修复。本公开提供了可用于促进骨生长的重组核酸。例如，本公开提供编码成纤维细胞生长因子-2（FGF-2）类似物的重组核酸。本公开还提供了掺入这些核酸的载体和细胞，以及由它们编码的FGF-2类似物。本公开还提供了骨髓移植的小鼠系统以及用于生产的方法以及使用该系统的方法。还提供诱导造血干细胞分化和/或诱导分化的方法，以及用于增强骨生长和/或伤口修复（例如，骨折修复）的方法。

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