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公开(公告)号：US20110065907A1
公开(公告)日：2011-03-17
申请号：US12914338
申请日：2010-10-28
申请人： Adrian Salic , Timothy J. Mitchison
发明人： Adrian Salic , Timothy J. Mitchison
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. In particular, the methods include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
摘要翻译： 本发明涉及体外和体内标记核酸聚合物的方法。 特别地，所述方法包括掺入核酸聚合物的核苷酸类似物与附着于标签的试剂之间的[3 + 2]环加成。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。
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公开(公告)号：US20100311063A1
公开(公告)日：2010-12-09
申请号：US12785999
申请日：2010-05-24
申请人： Kyle R. Gee , Brian Agnew , Adrian Salic , Timothy J. Mitchison
发明人： Kyle R. Gee , Brian Agnew , Adrian Salic , Timothy J. Mitchison
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. Certain methods are provided that include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Other methods are provided that include a Staudinger ligation between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent comprising a substituted triarylphosphine attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
摘要翻译： 本发明涉及体外和体内标记核酸聚合物的方法。 提供了包括掺入核酸聚合物的核苷酸类似物与附着于标记物上的试剂之间的[3 + 2]环加成的某些方法。 提供了包括掺入核酸聚合物的核苷酸类似物与包含连接到标签上的取代的三芳基膦的试剂之间的施陶丁格连接的其它方法。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。
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公开(公告)号：US07767421B2
公开(公告)日：2010-08-03
申请号：US11588697
申请日：2006-10-27
申请人： Kyle R. Gee , Brian Agnew , Adrian Salic , Timothy J. Mitchison
发明人： Kyle R. Gee , Brian Agnew , Adrian Salic , Timothy J. Mitchison
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. Certain methods are provided that include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Other methods are provided that include a Staudinger ligation between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent comprising a substituted triarylphosphine attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
摘要翻译： 本发明涉及体外和体内标记核酸聚合物的方法。 提供了包括掺入核酸聚合物的核苷酸类似物与附着于标记物上的试剂之间的[3 + 2]环加成的某些方法。 提供了包括掺入核酸聚合物的核苷酸类似物与包含连接到标签上的取代的三芳基膦的试剂之间的施陶丁格连接的其它方法。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。
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公开(公告)号：US08541570B2
公开(公告)日：2013-09-24
申请号：US12785999
申请日：2010-05-24
申请人： Kyle R. Gee , Brian Agnew , Adrian Salic , Timothy J. Mitchison
发明人： Kyle R. Gee , Brian Agnew , Adrian Salic , Timothy J. Mitchison
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. Certain methods are provided that include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Other methods are provided that include a Staudinger ligation between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent comprising a substituted triarylphosphine attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
摘要翻译： 本发明涉及体外和体内标记核酸聚合物的方法。 提供了包括掺入核酸聚合物的核苷酸类似物与附着于标记物上的试剂之间的[3 + 2]环加成的某些方法。 提供了包括掺入核酸聚合物的核苷酸类似物与包含连接到标签上的取代的三芳基膦的试剂之间的施陶丁格连接的其它方法。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。
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公开(公告)号：US08859753B2
公开(公告)日：2014-10-14
申请号：US12914338
申请日：2010-10-28
申请人： Adrian Salic , Timothy J. Mitchison
发明人： Adrian Salic , Timothy J. Mitchison
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. In particular, the methods include a [3+2] cycloaddition between a nucleotide analog incorporated into a nucleic acid polymer and a reagent attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
摘要翻译： 本发明涉及体外和体内标记核酸聚合物的方法。 特别地，所述方法包括掺入核酸聚合物的核苷酸类似物与附着于标签的试剂之间的[3 + 2]环加成。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。
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公开(公告)号：US07910335B2
公开(公告)日：2011-03-22
申请号：US11588732
申请日：2006-10-27
申请人： Adrian Salic , Timothy J. Mitchison
发明人： Adrian Salic , Timothy J. Mitchison
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. In particular, the methods include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
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公开(公告)号：US08987514B2
公开(公告)日：2015-03-24
申请号：US13147453
申请日：2010-02-04
申请人： Adrian Salic
发明人： Adrian Salic
IPC分类号： C12Q1/02 , C07C213/00 , C07C215/00 , C07C217/00 , C07C247/00 , A61K49/00 , A61K47/48 , G01N33/92 , G01N1/30
CPC分类号： A61K49/0041 , A61K47/4813 , A61K47/555 , G01N1/30 , G01N33/92
摘要： The present invention provides a method to label phospholipids in vivo based on the metabolic incorporation of an alkynyl- or azido-labeled metabolic precursor into phospholipids. The resulting phospholipids have alkynyl or azido moieties, which, upon reaction with a labeled azide or alkyne, respectively, form labeled compounds that can be visualized using optical or electron microscopy with high sensitivity and spatial resolution in cells or tissue. The present method provides a valuable tool for imaging phospholipid synthesis, turnover and subcellular localization in cultured cells as well as in animals.
摘要翻译： 本发明提供了一种基于将炔基或叠氮基标记的代谢前体代谢并入磷脂中而在体内标记磷脂的方法。 所得磷脂具有炔基或叠氮基部分，其分别与标记的叠氮化物或炔反应形成标记的化合物，其可以使用光学或电子显微镜在细胞或组织中具有高灵敏度和空间分辨率可视化。 本方法为培养细胞以及动物中磷脂合成，周转和亚细胞定位的成像提供了有价值的工具。
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公开(公告)号：US20120028290A1
公开(公告)日：2012-02-02
申请号：US13147453
申请日：2010-02-04
申请人： Adrian Salic
发明人： Adrian Salic
IPC分类号： C12Q1/02 , C07C247/06 , C07C247/12 , C07C215/24 , C07C247/04
CPC分类号： A61K49/0041 , A61K47/4813 , A61K47/555 , G01N1/30 , G01N33/92
摘要： The present invention provides a method to label phospholipids in vivo based on the metabolic incorporation of an alkynyl- or azido-labeled metabolic precursor into phospholipids. The resulting phospholipids have alkynyl or azido moieties, which, upon reaction with a labeled azide or alkyne, respectively, form labeled compounds that can be visualized using optical or electron microscopy with high sensitivity and spatial resolution in cells or tissue. The present method provides a valuable tool for imaging phospholipid synthesis, turnover and subcellular localization in cultured cells as well as in animals.
摘要翻译： 本发明提供了一种基于将炔基或叠氮基标记的代谢前体代谢并入磷脂中而在体内标记磷脂的方法。 所得磷脂具有炔基或叠氮基部分，其分别与标记的叠氮化物或炔反应形成标记的化合物，其可以使用光学或电子显微镜在细胞或组织中具有高灵敏度和空间分辨率可视化。 本方法为培养细胞以及动物中磷脂合成，周转和亚细胞定位的成像提供了有价值的工具。
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公开(公告)号：US20070207476A1
公开(公告)日：2007-09-06
申请号：US11588732
申请日：2006-10-27
申请人： Adrian Salic , Timothy Mitchison
发明人： Adrian Salic , Timothy Mitchison
CPC分类号： C12Q1/6806 , C12Q1/68 , C12Q1/6816 , C12Q1/6841 , G01N33/5011 , G01N33/582 , C12Q2525/101 , C12Q2563/107
摘要： The present invention relates to methods for the labeling of nucleic acid polymers in vitro and in vivo. In particular, the methods include a [3+2] cycloaddition between a nucleotide analogue incorporated into a nucleic acid polymer and a reagent attached to a label. Such methods do not require fixation and denaturation and therefore can be applied to the labeling of nucleic acid polymers in living cells and in organisms. Also provided are methods for measuring cellular proliferation. In these methods, the amount of label incorporated into the DNA is measured as an indication of cellular proliferation. The methods of the invention can be used in a wide variety of applications including clinical diagnosis of diseases and disorders in which cellular proliferation is involved, toxicity assays, and as a tool for the study of chromosomes' ultrastructures.
摘要翻译： 本发明涉及体外和体内标记核酸聚合物的方法。 特别地，所述方法包括掺入核酸聚合物的核苷酸类似物与附着于标签的试剂之间的[3 + 2]环加成。 这种方法不需要固定和变性，因此可以应用于活细胞和生物体中核酸聚合物的标记。 还提供了测量细胞增殖的方法。 在这些方法中，测量结合到DNA中的标记量作为细胞增殖的指示。 本发明的方法可用于各种应用，包括涉及细胞增殖的疾病和病症的临床诊断，毒性测定以及用于研究染色体超微结构的工具。
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