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1. 发明授权

US06271341B1 Transcriptional activators with graded transactivation potential 有权
标题翻译：具有分级反式激活潜能的转录激活子
公开(公告)号：US06271341B1
公开(公告)日：2001-08-07
申请号：US09577027
申请日：2000-05-23
申请人： Udo Baron , Manfred Gossen , Hermann Bujard
发明人： Udo Baron , Manfred Gossen , Hermann Bujard
IPC分类号： C07K1900
CPC分类号： C07K14/005 , A01K2217/05 , A61K48/00 , C07K2319/00 , C12N15/635 , C12N15/85 , C12N2710/16622 , C12N2830/003 , C12N2830/006
摘要： Transcriptional activators which differ in their activation potential by more than 3 orders of magnitude are provided. The transactivators are fusions between a DNA binding protein (e.g., a Tet repressor) and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). Substitution mutations at amino acid position 442 within the minimal VP16 domain provide transactivators with differing transactivation ability. Moreover, chimeric activation domains comprising both wild type and mutant minimal VP16 domains provide additional variants with differing transactivation ability. Various aspects of the invention pertain to nucleic acid molecules, vectors, host cells, fusion proteins, transgenic and homologous recombinant organisms and methods of regulating gene transcription.
摘要翻译：提供了其激活电位不同于3个数量级的转录激活子。反式激活因子是DNA结合蛋白（例如Tet阻遏物）和衍生自单纯疱疹病毒蛋白16（VP16）的最小转录激活结构域之间的融合物。在最小VP16结构域内的氨基酸位置442处的取代突变体提供具有不同反式激活能力的反式激活因子。此外，包含野生型和突变型最小VP16结构域的嵌合活化域提供了具有不同反式激活能力的另外的变体。本发明的各个方面涉及核酸分子，载体，宿主细胞，融合蛋白，转基因和同源重组生物以及调节基因转录的方法。

2. 发明授权

US5789156A Tetracycline-regulated transcriptional inhibitors 失效
标题翻译：四环素调节的转录抑制剂
公开(公告)号：US5789156A
公开(公告)日：1998-08-04
申请号：US383754
申请日：1995-02-03
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , C07K14/035 , C07K14/245 , C12N1/21 , C12N15/63 , C12N15/67 , C12N15/85 , C12Q1/68 , C07H21/04
CPC分类号： C12N15/8509 , C07K14/005 , C07K14/245 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , A01K2217/05 , A01K2217/075 , A01K2217/20 , A01K2267/01 , A01K2267/02 , A01K2267/03 , A01K2267/0393 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2710/16622 , C12N2830/003 , C12N2830/006 , Y10S435/81
摘要： Nucleic acid molecules and proteins useful for regulating the expression of genes in eukaryotic cells and organisms in a highly controlled manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is inhibited by a transcriptional inhibitor fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences either (i) in the absence but not the presence of tetracycline (or an analogue thereof) or (ii) in the presence but not the absence of tetracycline (or an analogue thereof), and a second polypeptide which directly or indirectly inhibits transcription in eukaryotic cells. In one embodiment, the fusion protein comprises a Tet repressor operatively linked to a transcriptional silencer polypeptide. In another embodiment, the fusion protein comprises a mutated Tet repressor operatively linked to a transcriptional silencer polypeptide. The fusion proteins of the invention are useful for reducing the level of transcription of a tet operator-linked target gene. Moreover, the fusion proteins of the invention can be used in combination with tetracycline-regulated transcriptional activator fusion proteins to allow for precise regulation of the expression of one or multiple target genes. Kits including the components of the regulatory system of the invention are also encompassed by the invention.
摘要翻译：公开了用于以高度受控的方式调节真核细胞和生物体中基因表达的核酸分子和蛋白质。在本发明的调节系统中，tet操纵子连接的核苷酸序列的转录受到由两个多肽组成的转录抑制剂融合蛋白的抑制，所述两个多肽是结合tet操纵子序列的第一个多肽，（i）在不存在但不存在的情况下的四环素（或其类似物）或（ii）在存在但不存在四环素（或其类似物）的情况下，以及在真核细胞中直接或间接抑制转录的第二多肽。在一个实施方案中，融合蛋白包含可操作地连接到转录沉默子多肽的Tet阻遏物。在另一个实施方案中，融合蛋白包含与转录沉默子多肽有效连接的突变Tet阻遏物。本发明的融合蛋白可用于降低与tet操纵子连接的靶基因的转录水平。此外，本发明的融合蛋白可以与四环素调节的转录激活物融合蛋白组合使用，以允许精确调节一个或多个靶基因的表达。包括本发明的调节系统的组分的试剂盒也包括在本发明中。

3. 发明授权

US5654168A Tetracycline-inducible transcriptional activator and tetracycline-regulated transcription units 失效
标题翻译：四环素诱导型转录激活因子和四环素调节转录单位
公开(公告)号：US5654168A
公开(公告)日：1997-08-05
申请号：US275876
申请日：1994-07-15
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , C07K14/245 , C12N15/63 , C12N15/67 , C12N15/85 , C12P21/00 , C07H21/04 , C12N15/00
CPC分类号： C12N15/8509 , C07K14/245 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , A01K2217/05 , A01K2217/20 , A01K2267/01 , A01K2267/03 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2830/003 , C12N2830/006
摘要： Nucleic acid molecules and proteins useful for regulating the expression of genes in eukaryotic cells and organisms in an inducible manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is stimulated by a transcriptional activator fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences in the presence of tetracycline or a tetracycline analogue and a second polypeptide which directly or indirectly activates transcription in eukaryotic cells. In one embodiment, the fusion protein comprises a mutated Tet repressor operatively linked to a transcriptional activation polypeptide, such as a portion of herpes simplex virus virion protein 16. In the absence of an inducing agent (tetracycline or a tetracycline analogue), transcription of the tet operator-linked nucleotide sequence remains uninduced. In the presence of the inducing agent, transcription of the tet operator-linked nucleotide sequence is stimulated by the transactivator fusion protein of the invention. Novel transcription units which allow for coordinate or independent tetracycline-regulated expression of two or more nucleotide sequences by the transactivator fusion protein of the invention are also disclosed. Kits including the components of the regulatory system of the invention are also encompassed by the invention.
摘要翻译：公开了用于以可诱导的方式调节真核细胞和生物体中基因表达的核酸分子和蛋白质。在本发明的调节系统中，tet操纵子连接的核苷酸序列的转录受到由两个多肽组成的转录激活物融合蛋白的刺激，该多肽是在四环素或四环素类似物存在下与tet操纵子序列结合的第一多肽在真核细胞中直接或间接激活转录的第二多肽。在一个实施方案中，融合蛋白包含可操作地连接到转录激活多肽（例如一部分单纯疱疹病毒颗粒蛋白16）的突变的Tet阻遏物。在不存在诱导剂（四环素或四环素类似物）的情况下， tet操纵子连接的核苷酸序列仍未引起。在诱导剂存在下，通过本发明的反式激活因子融合蛋白刺激tet操纵子连接的核苷酸序列的转录。还公开了通过本发明的反式激活物融合蛋白允许两个或更多个核苷酸序列的协调或独立的四环素调节的表达的新型转录单位。包括本发明的调节系统的组分的试剂盒也包括在本发明中。

4. 发明申请

US20110217367A1 MEANS FOR INHIBITING THE EXPRESSION OF ORC-1 失效
标题翻译：抑制ORC-1表达的方法
公开(公告)号：US20110217367A1
公开(公告)日：2011-09-08
申请号：US13120430
申请日：2009-09-23
申请人： Manfred Gossen , Ansgar Santel , Jorg Kaufmann
发明人： Manfred Gossen , Ansgar Santel , Jorg Kaufmann
IPC分类号： A61K9/127 , C12N15/63 , C12N5/10 , C07H21/00 , A61K31/713 , C12N5/071 , A61P35/02 , A61P31/22 , A61P35/00 , A61P31/20
CPC分类号： C12N15/1135 , C12N15/113 , C12N2310/14
摘要： The present invention is related to a nucleic acid molecule comprising a double-stranded structure, whereby the double-stranded structure comprises a first strand and a second strand, whereby the first strand comprises a first stretch of contiguous nucleotides and said first stretch is at least partially complementary to a target nucleic acid, and whereby the second strand comprises a second stretch of contiguous nucleotides and said second stretch is at least partially complementary to the first stretch, whereby the first stretch comprises a nucleic acid sequence which is at least complementary to a nucleotide core sequence of the nucleic acid sequence according to SEQ ID NO: 1, whereby the nucleotide core sequence comprises the nucleotide sequence from nucleotide positions 1755 to 1763 of SEQ ID NO: 1; from nucleotide positions 1904 to 1912 of SEQ. ID. No.1; from nucleotide positions 1905 to 1913 of SEQ ID NO: 1; from nucleotide positions 2548 to 2556 of SEQ ID NO: 1; whereby the first stretch is additionally at least partially complementary to a region preceding the 5′ end of the nucleotide core sequence and/or to a region following the 3′ end of the nucleotide core sequence.
摘要翻译：本发明涉及包含双链结构的核酸分子，由此双链结构包含第一链和第二链，其中第一链包含连续核苷酸的第一段，并且所述第一段至少是与靶核酸部分互补，并且其中所述第二链包含连续核苷酸的第二段，所述第二段与所述第一段至少部分互补，其中所述第一段包含与至少互补的核酸序列核苷酸核心序列包含SEQ ID NO：1的核苷酸序列1755至1763的核苷酸序列; 来自SEQ ID NO：1的核苷酸位置1904至1912。 ID。第一; 从SEQ ID NO：1的核苷酸位置1905到1913; 从SEQ ID NO：1的核苷酸位置2548至2556; 其中第一次拉伸另外至少部分地与核苷酸核心序列的5'末端之前的区域和/或与核苷酸核心序列的3'末端之后的区域互补。

5. 发明授权

US06271348B1 Tetracycline-inducible transcriptional inhibitor fusion proteins 有权
标题翻译：四环素诱导型转录抑制剂融合蛋白
公开(公告)号：US06271348B1
公开(公告)日：2001-08-07
申请号：US09489777
申请日：2000-01-24
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： C07K1900
CPC分类号： C07K14/005 , A01K67/0275 , A01K2217/05 , A01K2217/075 , A01K2217/20 , A01K2227/105 , A01K2267/0393 , A61K48/00 , C07K14/245 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N15/62 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , C12N15/8509 , C12N2710/16622 , C12N2830/003 , C12N2830/006 , C12N2830/32
摘要： Methods of regulating gene expression in subjects using tetracycline-responsive fusion proteins are disclosed. In one embodiment, the method involves introducing into a cell the subject a nucleic acid molecule encoding a fusion protein which inhibits transcription, the fusion protein comprising a first polypeptide which binds to a tet operator sequence, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first polypeptide can binds to a tet operator sequence in the absence, but not the presence, of tetracycline. Alternatively, the first polypeptide can binds to a tet operator sequence in the presence, but not the absence, of tetracycline. In another embodiment, the method of the invention involves obtaining a cell from a subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding an inhibitory fusion protein of the invention to form a modified cell, administering the modified cell to the subject and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecules can be linked or can be separate molecules.
摘要翻译：公开了使用四环素响应融合蛋白调节受试者基因表达的方法。在一个实施方案中，该方法包括向受试者引入编码抑制转录的融合蛋白的核酸分子，该融合蛋白包含与tet操纵子序列结合的第一多肽，该第一多肽与抑制转录的异源第二多肽有效连接在真核细胞中并调节受试者中四环素或其类似物的浓度。在不存在但不存在四环素的情况下，第一种多肽可以结合tet操纵子序列。或者，第一多肽可以在四环素存在下但不存在时与tet操纵子序列结合。在另一个实施方案中，本发明的方法涉及从受试者获得细胞，将在第一核酸分子中引入至少一个tet操纵子序列的第一核酸分子，将编码本发明的抑制性融合蛋白以形成修饰的细胞，向受试者施用修饰的细胞并调节受试者中四环素或其类似物的浓度。第一和第二核酸分子可以连接或可以是分开的分子。

6. 发明授权

US08722875B2 Means for inhibiting the expression of Orc-1 失效
标题翻译：抑制Orc-1表达的方法
公开(公告)号：US08722875B2
公开(公告)日：2014-05-13
申请号：US13120430
申请日：2009-09-23
申请人： Manfred Gossen , Ansgar Santel , Jorg Kaufmann
发明人： Manfred Gossen , Ansgar Santel , Jorg Kaufmann
IPC分类号： C07H21/04 , C12Q1/68 , C12N15/11 , C12N5/02 , C12N5/00
CPC分类号： C12N15/1135 , C12N15/113 , C12N2310/14
摘要： The present invention is related to a nucleic acid molecule comprising a double-stranded structure, whereby the double-stranded structure comprises a first strand and a second strand, whereby the first strand comprises a first stretch of contiguous nucleotides and said first stretch is at least partially complementary to a target nucleic acid, and whereby the second strand comprises a second stretch of contiguous nucleotides and said second stretch is at least partially complementary to the first stretch, whereby the first stretch comprises a nucleic acid sequence which is at least complementary to a nucleotide core sequence of the nucleic acid sequence according to SEQ ID NO: 1, whereby the nucleotide core sequence comprises the nucleotide sequence from nucleotide positions 1755 to 1763 of SEQ ID NO: 1; from nucleotide positions 1904 to 1912 of SEQ.ID.No.1; from nucleotide positions 1905 to 1913 of SEQ ID NO: 1; from nucleotide positions 2548 to 2556 of SEQ ID NO: 1; whereby the first stretch is additionally at least partially complementary to a region preceding the 5′ end of the nucleotide core sequence and/or to a region following the 3′ end of the nucleotide core sequence.
摘要翻译：本发明涉及包含双链结构的核酸分子，由此双链结构包含第一链和第二链，其中第一链包含连续核苷酸的第一段，并且所述第一段至少是与靶核酸部分互补，并且其中所述第二链包含连续核苷酸的第二段，所述第二段与所述第一段至少部分互补，由此所述第一段包含与至少互补的核酸序列核苷酸核心序列包含SEQ ID NO：1的核苷酸序列1755至1763的核苷酸序列; 从SEQ.ID.No.1的核苷酸位置1904到1912; 从SEQ ID NO：1的核苷酸位置1905到1913; 从SEQ ID NO：1的核苷酸位置2548至2556; 其中第一次拉伸另外至少部分地与核苷酸核心序列的5'末端之前的区域和/或与核苷酸核心序列的3'末端之后的区域互补。

7. 发明授权

US6136954A Tetracycline-inducible transcriptional activator fusion proteins 有权
标题翻译：四环素诱导型转录激活融合蛋白
公开(公告)号：US6136954A
公开(公告)日：2000-10-24
申请号：US162184
申请日：1998-09-28
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , A61K48/00 , C07K14/00 , C07K14/035 , C07K14/195 , C07K14/245 , C07K19/00 , C12N15/11 , C12N15/63 , C12N15/67 , C12N15/85
CPC分类号： C07K14/005 , A01K67/0275 , C07K14/245 , C12N15/62 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , C12N15/8509 , A01K2217/05 , A01K2217/075 , A01K2217/20 , A01K2227/105 , A01K2267/0393 , A61K48/00 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2710/16622 , C12N2830/003 , C12N2830/006 , C12N2830/32
摘要： Methods of regulating gene expression in subjects using tetracycline-responsive fusion proteins are disclosed. In one embodiment, the method involves introducing into a cell the subject a nucleic acid molecule encoding a fusion protein which inhibits transcription, the fusion protein comprising a first polypeptide which binds to a tet operator sequence, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first polypeptide can binds to a tet operator sequence in the absence, but not the presence, of tetracycline. Alternatively, the first polypeptide can binds to a tet operator sequence in the presence, but not the absence, of tetracycline. In another embodiment, the method of the invention involves obtaining a cell from a subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding an inhibitory fusion protein of the invention to form a modified cell, administering the modified cell to the subject and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecules can be linked or can be separate molecules.
摘要翻译：公开了使用四环素响应融合蛋白调节受试者基因表达的方法。在一个实施方案中，该方法包括向受试者引入编码抑制转录的融合蛋白的核酸分子，该融合蛋白包含与tet操纵子序列结合的第一多肽，该第一多肽与抑制转录的异源第二多肽有效连接在真核细胞中并调节受试者中四环素或其类似物的浓度。在不存在但不存在四环素的情况下，第一种多肽可以结合tet操纵子序列。或者，第一多肽可以在四环素存在下但不存在时与tet操纵子序列结合。在另一个实施方案中，本发明的方法涉及从受试者获得细胞，将在第一核酸分子中引入至少一个tet操纵子序列的第一核酸分子，将编码本发明的抑制性融合蛋白以形成修饰的细胞，向受试者施用修饰的细胞并调节受试者中四环素或其类似物的浓度。第一和第二核酸分子可以连接或可以是分开的分子。

8. 发明授权

US6004941A Methods for regulating gene expression 失效
标题翻译：调节基因表达的方法
公开(公告)号：US6004941A
公开(公告)日：1999-12-21
申请号：US485740
申请日：1995-06-07
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , A61K48/00 , C07K14/00 , C07K14/035 , C07K14/195 , C07K14/245 , C07K19/00 , C12N15/11 , C12N15/63 , C12N15/67 , C12N15/85
CPC分类号： C07K14/005 , A01K67/0275 , C07K14/245 , C12N15/63 , C12N15/635 , C12N15/67 , C12N15/85 , C12N15/8509 , A01K2217/05 , A01K2217/075 , A01K2217/20 , A01K2227/105 , A01K2267/01 , A01K2267/02 , A01K2267/03 , A01K2267/0393 , A61K48/00 , C07K2319/00 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2710/16622 , C12N2830/003 , C12N2830/006 , C12N2830/32
摘要： Methods of regulating gene expression in subjects using tetracycline-responsive fusion proteins are disclosed. In one embodiment, the method involves introducing into a cell the subject a nucleic acid molecule encoding a fusion protein which activates transcription, the fusion protein comprising a first polypeptide which binds to a tet operator sequence in the presence of tetracycline or a tetracycline analogue operatively linked to a second polypeptide which activates transcription in eukaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. In another embodiment, the cell further comprises a fusion protein which inhibits transcription, the fusion protein comprising a first polypeptide which binds to a tet operator sequence, operatively linked to a second polypeptide which inhibits transcription in eukaryotic cells. In yet another embodiment, the method involves obtaining a cell from a subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding a fusion protein of the invention to form a modified cell, administering the modified cell to the subject and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecules can be linked or can be separate molecules.
摘要翻译：公开了使用四环素响应融合蛋白调节受试者基因表达的方法。在一个实施方案中，该方法包括向受试者引入编码活化转录的融合蛋白的核酸分子，该融合蛋白包含在四环素或四环素类似物的存在下与tet操纵子序列结合的第一多肽，所述四环素或四环素类似物可操作地连接涉及在真核细胞中激活转录的第二种多肽; 并调节受试者中四环素或其类似物的浓度。在另一个实施方案中，所述细胞还包含抑制转录的融合蛋白，所述融合蛋白包含结合tet操纵子序列的第一多肽，所述第一多肽与抑制真核细胞转录的第二多肽有效连接。在另一个实施方案中，该方法包括从受试者获得细胞，将在第一核酸分子中引入至少一个tet操纵子序列的第一核酸分子，将编码融合蛋白的第二核酸分子以形成修饰的细胞，将修饰的细胞给予受试者并调节受试者中四环素或其类似物的浓度。第一和第二核酸分子可以连接或可以是分开的分子。

9. 发明授权

US5888981A Methods for regulating gene expression 失效
标题翻译：调节基因表达的方法
公开(公告)号：US5888981A
公开(公告)日：1999-03-30
申请号：US479306
申请日：1995-06-07
申请人： Hermann Bujard , Manfred Gossen , Jochen G. Salfeld , Jeffrey W. Voss
发明人： Hermann Bujard , Manfred Gossen , Jochen G. Salfeld , Jeffrey W. Voss
IPC分类号： A01K67/027 , A61K48/00 , C12N5/08 , C12N15/62 , C12N15/63 , C12N15/85 , C12N15/00
CPC分类号： C12N15/62 , A01K67/0275 , C12N15/635 , C12N15/85 , A01K2217/05 , A01K2217/075 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N2830/006
摘要： A method for regulating expression of a tet operator-linked gene in a cell of a subject is disclosed. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a tetracycline-controllable transactivator (tTA), the tTA comprising a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. Alternatively, in another embodiment, the method involves obtaining the cell from the subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding a tTA, to form a modified cell, administering the modified cell to the is subject, and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecule can be within a single molecule (e.g., in the same vector) or on separate molecules.
摘要翻译：公开了一种用于调节受试者细胞中tet操纵子连接基因表达的方法。在一个实施方案中，所述方法包括将编码四环素可控反式激活因子（tTA）的核酸分子引入细胞，所述tTA包含可操作地连接到直接或间接激活真核细胞中的转录的多肽的Tet阻遏物; 并调节受试者中四环素或其类似物的浓度。或者，在另一个实施方案中，所述方法包括从受试者获得细胞，向细胞中引入第一核酸分子，该第一核酸分子可操作地将基因连接至至少一个tet操纵子序列，将编码tTA的第二核酸分子，以形成修饰的细胞，将修饰的细胞给予受试者，并调节受试者中四环素或其类似物的浓度。第一和第二核酸分子可以在单分子内（例如，在同一载体中）或分开的分子上。

10. 发明授权

US06914124B2 Tetracycline-regulated transcriptional activator fusion proteins 失效
标题翻译：四环素调节的转录激活融合蛋白
公开(公告)号：US06914124B2
公开(公告)日：2005-07-05
申请号：US09777317
申请日：2001-02-05
申请人： Hermann Bujard , Manfred Gossen
发明人： Hermann Bujard , Manfred Gossen
IPC分类号： A01K67/027 , A61K48/00 , C07K14/00 , C12N5/08 , C12N5/10 , C12N15/00 , C12N15/09 , C12N15/31 , C12N15/62 , C12N15/63 , C12N15/85 , C12R1/91
CPC分类号： A01K67/0275 , A01K2217/05 , A01K2217/075 , C07K2319/09 , C07K2319/61 , C07K2319/71 , C07K2319/80 , C12N15/62 , C12N15/635 , C12N15/85 , C12N2830/006
摘要： A method for regulating expression of a tet operator-linked gene in a cell of a subject is disclosed. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a tetracycline-controllable transactivator (tTA), the tTA comprising a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. Alternatively, in another embodiment, the method involves obtaining the cell from the subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding a tTA, to form a modified cell, administering the modified cell to the subject, and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecule can be within a single molecule (e.g., in the same vector) or on separate molecules.
摘要翻译：公开了一种用于调节受试者细胞中tet操纵子连接基因表达的方法。在一个实施方案中，所述方法包括将编码四环素可控反式激活因子（tTA）的核酸分子引入细胞，所述tTA包含可操作地连接到直接或间接激活真核细胞中的转录的多肽的Tet阻遏物; 并调节受试者中四环素或其类似物的浓度。或者，在另一个实施方案中，所述方法包括从受试者获得细胞，向细胞中引入第一核酸分子，该第一核酸分子可操作地将基因连接至至少一个tet操纵子序列，将编码tTA的第二核酸分子，以形成修饰的细胞，向受试者施用修饰的细胞，并调节受试者中四环素或其类似物的浓度。第一和第二核酸分子可以在单分子内（例如，在同一载体中）或分开的分子上。

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