公开(公告)号：US20070155014A1

公开(公告)日：2007-07-05

申请号：US11490474

申请日：2006-07-20

申请人： Luciana Bertolini ,  Michaeline Bunting ,  Knut Madden ,  Elizabeth Maga ,  James Murray

发明人： Luciana Bertolini ,  Michaeline Bunting ,  Knut Madden ,  Elizabeth Maga ,  James Murray

IPC分类号： C12N15/09 ,  C12N5/10

CPC分类号： C12N15/113 ,  C12N15/902 ,  C12N2310/11 ,  C12N2310/14

摘要： Methods and compositions for increasing or decreasing homologous recombination activity in a eukaryotic cell are provided. More particularly, various methods and compositions are provided which decrease the level of non-homologous recombination in a cell and thereby increase the frequency of targeted homologous recombination events. Various compositions including cells and kits that can be employed in the methods are also provided.

摘要翻译： 提供了用于增加或减少真核细胞中的同源重组活性的方法和组合物。 更具体地，提供了降低细胞中非同源重组水平从而增加靶向同源重组事件的频率的各种方法和组合物。 还提供了可用于该方法的各种组合物，包括可用于该方法的细胞和试剂盒。

公开(公告)号：US20060078902A1

公开(公告)日：2006-04-13

申请号：US11105594

申请日：2005-04-14

申请人： Michaeline Bunting ,  Knut Madden ,  Adam Harris

发明人： Michaeline Bunting ,  Knut Madden ,  Adam Harris

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6897 ,  C12N15/111 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2320/11 ,  C12N2330/30

摘要： The invention provides methods and compositions related to the field of gene expression regulation. In particular, methods and compositions of the invention can be used to identify RNAi cleavage sites along a target RNA molecule. Methods and compositions of the invention may also be used to knockdown expression of nucleic acid molecules which encode reporters.

摘要翻译： 本发明提供了与基因表达调控领域相关的方法和组合。 特别地，本发明的方法和组合物可用于鉴定沿靶RNA分子的RNAi切割位点。 本发明的方法和组合物也可用于敲低编码报道子的核酸分子的表达。

公开(公告)号：US20070196838A1

公开(公告)日：2007-08-23

申请号：US11612957

申请日：2006-12-19

申请人： Jonathan Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gray Temple

发明人： Jonathan Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James Hartley ,  Devon Byrd ,  Gray Temple

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P21/06 ,  C12N9/22

CPC分类号： C12N9/90 ,  C12P19/34

摘要： The present invention provides compositions and methods for recombinational cloning. The compositions vectors having multiple recombination sites and/or multiple topoisomerase recognition sities. The methods premit the simultaeous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 具有多个重组位点和/或多个拓扑异构酶识别性质的组合物载体。 该方法预先提出了两种或多种不同核酸分子的同源克隆。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供了包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。

公开(公告)号：US20050282184A1

公开(公告)日：2005-12-22

申请号：US11053187

申请日：2005-02-07

申请人： Jonathan Chesnut ,  Stewart Shuman ,  Knut Madden ,  John Heyman ,  Robert Bennett

发明人： Jonathan Chesnut ,  Stewart Shuman ,  Knut Madden ,  John Heyman ,  Robert Bennett

IPC分类号： A01K67/027 ,  C12N1/21 ,  C12N5/10 ,  C12N15/09 ,  C12N15/10 ,  C12N15/64 ,  C12N15/66 ,  C40B40/02 ,  C40B50/06 ,  C12Q1/68 ,  C12P19/34

CPC分类号： C12N15/66 ,  C12N15/10 ,  C12N15/64

摘要： The present invention provides compositions, methods, and kits for covalently linking nucleic acid molecules. The methods include a strand invasion step, and the compositions and kits are useful for performing such methods. For example, a method of covalently linking double stranded (ds) nucleic acid molecules can include contacting a first ds nucleic acid molecule, which has a topoisomerase linked to a 3′ terminus of one end and has a single stranded 5′ overhang at the same end, with a second ds nucleic acid molecule having a blunt end, such that the 5′ overhang can hybridize to a complementary sequence of the blunt end of the second nucleic acid molecule, and the topoisomerase can covalently link the ds nucleic acid molecules. The methods are simpler and more efficient than previous methods for covalently linking nucleic acid sequences, and the compositions and kits facilitate practicing the methods, including methods of directionally linking two or more ds nucleic acid molecules.

公开(公告)号：US20050069929A1

公开(公告)日：2005-03-31

申请号：US10913501

申请日：2004-08-09

申请人： Jonathan Chestnut ,  Knut Madden ,  Miroslav Dudas ,  Louis Leong ,  Adam Harris

发明人： Jonathan Chestnut ,  Knut Madden ,  Miroslav Dudas ,  Louis Leong ,  Adam Harris

IPC分类号： A61K48/00 ,  C07H21/04 ,  C12N20060101 ,  C12N15/10 ,  C12N15/11 ,  C12N15/64 ,  C12N15/66 ,  C12N15/867 ,  C12P19/34 ,  C12Q1/68

CPC分类号： C12P19/34 ,  C07H21/04 ,  C12N9/90 ,  C12N15/10 ,  C12N15/111 ,  C12N15/64 ,  C12N15/66 ,  C12N15/86 ,  C12N2310/111 ,  C12N2310/14 ,  C12N2330/30 ,  C12N2740/16043 ,  C12N2800/30 ,  C12N2800/70 ,  C12Y599/01

摘要： The present invention is in the fields of biotechnology and molecular biology. More particularly, the present invention relates to cloning or subcloning one or more nucleic acid molecules comprising one or more type IIs restriction enzyme recognition sites. The present invention also embodies cloning such nucleic acid molecules using recombinational cloning methods such as those employing recombination sites and recombination proteins. The present invention also relates to nucleic acid molecules (including RNA and iRNA), as well as proteins, expressed from host cells produced using the methods of the present invention.

摘要翻译： 本发明涉及生物技术和分子生物学领域。 更具体地说，本发明涉及克隆或亚克隆一个或多个包含一个或多个II型限制酶识别位点的核酸分子。 本发明还使用重组克隆方法（例如使用重组位点和重组蛋白的那些）来体现克隆这种核酸分子。 本发明还涉及由使用本发明的方法制备的宿主细胞表达的核酸分子（包括RNA和iRNA）以及蛋白质。

公开(公告)号：US08030066B2

公开(公告)日：2011-10-04

申请号：US11612445

申请日：2006-12-18

申请人： Jonathan D. Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James L. Hartley ,  Devon R. Byrd ,  Gray F. Temple

发明人： Jonathan D. Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin Gleeson ,  James Fan ,  Michael Brasch ,  David Cheo ,  James L. Hartley ,  Devon R. Byrd ,  Gray F. Temple

IPC分类号： C12N15/00

CPC分类号： C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

摘要： The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供了包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。

公开(公告)号：US20080241889A1

公开(公告)日：2008-10-02

申请号：US11612445

申请日：2006-12-18

申请人： Jonathan D. Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin A.G. Gleeson ,  James Fan ,  Michael A. Brasch ,  David Cheo ,  James L. Hartley ,  Devon R.N. Byrd ,  Gray F. Temple

发明人： Jonathan D. Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin A.G. Gleeson ,  James Fan ,  Michael A. Brasch ,  David Cheo ,  James L. Hartley ,  Devon R.N. Byrd ,  Gray F. Temple

IPC分类号： C12P19/34 ,  C12N15/11 ,  C12N15/00 ,  C12N5/06

CPC分类号： C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

摘要： The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供了包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。

公开(公告)号：US07198924B2

公开(公告)日：2007-04-03

申请号：US10792035

申请日：2004-03-04

申请人： Jonathan D. Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin A. G. Gleeson ,  James Fan ,  Michael A. Brasch ,  David Cheo ,  James L. Hartley ,  Devon R. N. Byrd ,  Gary F. Temple

发明人： Jonathan D. Chesnut ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin A. G. Gleeson ,  James Fan ,  Michael A. Brasch ,  David Cheo ,  James L. Hartley ,  Devon R. N. Byrd ,  Gary F. Temple

IPC分类号： C12P19/34 ,  C12Q1/68

CPC分类号： C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

摘要： The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供了包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。

公开(公告)号：US20050054847A1

公开(公告)日：2005-03-10

申请号：US10902704

申请日：2004-07-30

申请人： Knut Madden ,  Adam Harris ,  Karl Hecker ,  Byung-in Lee

发明人： Knut Madden ,  Adam Harris ,  Karl Hecker ,  Byung-in Lee

IPC分类号： C07H21/02 ,  C12N20060101 ,  C12N15/11

CPC分类号： C07H21/02 ,  B01D15/3804 ,  C12N15/111 ,  C12N2330/31

摘要： The invention provides methods of preparing nucleic acids, such as RNA molecules, of a defined size or range of sizes. The invention provides compositions, methods and kits for use in the production and preparation of small RNA molecules (including without limitation micro-RNA, siRNA, d-siRNA and e-siRNA) and other nucleic acids of various sizes.

摘要翻译： 本发明提供了制备具有规定尺寸或尺寸范围的核酸如RNA分子的方法。 本发明提供了用于生产和制备小RNA分子（包括但不限于微RNA，siRNA，d-siRNA和e-siRNA）和各种尺寸的其它核酸的组合物，方法和试剂盒。

公开(公告)号：US20130004996A1

公开(公告)日：2013-01-03

申请号：US13219440

申请日：2011-08-26

申请人： Jonathan D. CHESNUT ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin A. G. Gleeson ,  James Fan ,  Michael A. Brasch ,  David Cheo ,  James L. Hartley ,  Devon R.N. Byrd ,  Gary F. Temple

发明人： Jonathan D. CHESNUT ,  John Carrino ,  Louis Leong ,  Knut Madden ,  Martin A. G. Gleeson ,  James Fan ,  Michael A. Brasch ,  David Cheo ,  James L. Hartley ,  Devon R.N. Byrd ,  Gary F. Temple

IPC分类号： C12P19/34 ,  C12N9/96 ,  C12N15/66 ,  C07H21/04

CPC分类号： C12N15/66 ,  C12N9/90 ,  C12N15/10 ,  C12N15/64 ,  C12N15/902 ,  C12N2800/108 ,  C12N2800/30 ,  C12N2800/70 ,  C12N2840/20 ,  C12P19/34 ,  C12Y599/01

摘要： The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according tb the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

摘要翻译： 本发明提供了用于重组克隆的组合物和方法。 组合物包括具有多个重组位点和/或多个拓扑异构酶识别位点的载体。 该方法允许同时克隆两种或更多种不同的核酸分子。 在一些实施方案中，分子融合在一起，而在其它实施方案中，分子被插入载体中的不同位点。 本发明还通常提供通过重组连接或连接许多可以相同或不同的分子和/或化合物（例如，化合物，药物，蛋白质或肽，脂质，核酸，碳水化合物等）。 本发明还提供包含本发明的核酸分子或根据本发明的方法制备的宿主细胞，并且还提供了包含本发明的组合物，宿主细胞和核酸分子的试剂盒，其可用于合成核酸分子 根据本发明的方法。