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    HYBRIDIZATION AND MISMATCH DISCRIMINATION USING OLIGONUCLEOTIDES CONJUGATED TO MINOR GROOVE BINDERS
    11.
    发明申请
    HYBRIDIZATION AND MISMATCH DISCRIMINATION USING OLIGONUCLEOTIDES CONJUGATED TO MINOR GROOVE BINDERS 失效
    使用与少部分粘结剂相结合的寡核苷酸进行杂交和错配歧视

    公开(公告)号：US20110275070A1

    公开(公告)日：2011-11-10

    申请号：US12847148

    申请日：2010-08-31

    申请人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, JR.

    发明人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, JR.

    IPC分类号： C12Q1/68

    CPC分类号： C07H21/00 ,  C07D207/34 ,  C07D403/12 ,  C07D487/04 ,  C07H19/073 ,  C07H19/173 ,  C07H19/23 ,  C07H21/04 ,  C12Q1/6813 ,  C12Q2527/107 ,  C12Q2525/197

    摘要： Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

    摘要翻译： 小沟结合分子（例如1,2-二氢 - （3H） - 吡咯并[3,2-e]吲哚-7-羧酸酯（CDPI3）的三聚体和寡核苷酸之间的缀合物形成非常稳定的具有互补靶的杂交体 序列，其中系留的CDPI3基团位于双链体的小沟槽中。 这些缀合物可以用作探针和引物。 由于其非常高的结合亲和力，短至8聚体的缀合物可以用作高特异性和高效的扩增引物。 MGB缀合还增加了短寡核苷酸的鉴别能力，提供了通过短寡核苷酸增强核苷酸序列错配检测。 本文所述的MGB缀合的探针和引物有助于各种分析和诊断程序，如扩增反应，PCR，单核苷酸多态性检测，基因敲除，差异显示，荧光能量转移，水解性探针测定等。 通过允许使用具有更高特异性和更好的辨别力的较短的寡核苷酸。

    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
    12.
    发明授权
    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders 失效
    使用与小沟结合物缀合的寡核苷酸的杂交和错配鉴别

    公开(公告)号：US06884584B2

    公开(公告)日：2005-04-26

    申请号：US10127936

    申请日：2002-04-22

    申请人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    发明人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    IPC分类号： C12N15/09 ,  C07H19/23 ,  C07H21/00 ,  C07H21/02 ,  C07H21/04 ,  C12Q1/68 ,  G01N21/78 ,  G01N33/483 ,  G01N33/53 ,  G01N33/566 ,  C07H19/04 ,  C12P19/34

    CPC分类号： C07H21/00 ,  C07H19/23 ,  C12Q1/6813 ,  C12Q2527/107 ,  C12Q2525/197

    摘要： Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

    摘要翻译： 小沟结合分子如1,2-二氢 - （3H） - 吡咯并[3,2-e]吲哚-7-羧酸酯（CDPI 3 N 3）的三聚体和 寡核苷酸形成具有互补靶序列的异常稳定的杂交体，其中连接的CDPI 3 基团位于双链体的小沟槽中。 这些缀合物可以用作探针和引物。 由于其非常高的结合亲和力，短至8聚体的缀合物可以用作高特异性和高效的扩增引物。 MGB缀合还增加了短寡核苷酸的鉴别能力，提供了通过短寡核苷酸增强核苷酸序列错配检测。 本文所述的MGB缀合的探针和引物有助于各种分析和诊断程序，如扩增反应，PCR，单核苷酸多态性检测，基因敲除，差异显示，荧光能量转移，水解性探针测定等。 通过允许使用具有更高特异性和更好的辨别力的较短的寡核苷酸。

    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders
    13.
    发明授权
    Hybridization and mismatch discrimination using oligonucleotides conjugated to minor groove binders 失效
    使用与小沟结合物缀合的寡核苷酸的杂交和错配鉴别

    公开(公告)号：US06312894B1

    公开(公告)日：2001-11-06

    申请号：US09054832

    申请日：1998-04-03

    申请人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    发明人： Joel Hedgpeth ,  Irina A. Afonina ,  Igor V. Kutyavin ,  Eugeny A. Lukhtanov ,  Evgeniy S. Belousov ,  Rich B. Meyer, Jr.

    IPC分类号： C12Q168

    CPC分类号： C07H21/00 ,  C07H19/23 ,  C12Q1/6813 ,  C12Q2527/107 ,  C12Q2525/197

    摘要： Conjugates between a minor groove binding molecule, such as the trimer of 1,2-dihydro-(3H)-pyrrolo[3,2-e]indole-7-carboxylate (CDPI3), and an oligonucleotide form unusually stable hybrids with complementary target sequences, in which the tethered CDPI3 group resides in the minor groove of the duplex. These conjugates can be used as probes and primers. Due to their unusually high binding affinity, conjugates as short as 8-mers can be used as amplification primers with high specificity and efficiency. MGB conjugation also increases the discriminatory power of short oligonucleotides, providing enhanced detection of nucleotide sequence mismatches by short oligonucleotides. The MGB-conjugated probes and primers described herein facilitate various analytic and diagnostic procedures, such as amplification reactions, PCR, detection of single-nucleotide polymorphisms, gene hunting, differential display, fluorescence energy transfer, hydrolyzable probe assays and others; by allowing the use of shorter oligonucleotides, which have higher specificity and better discriminatory power.

    摘要翻译： 小沟结合分子（例如1,2-二氢 - （3H） - 吡咯并[3,2-e]吲哚-7-羧酸酯（CDPI3）的三聚体和寡核苷酸之间的缀合物形成非常稳定的具有互补靶的杂交体 序列，其中系留的CDPI3基团位于双链体的小沟槽中。 这些缀合物可以用作探针和引物。 由于其非常高的结合亲和力，短至8聚体的缀合物可以用作高特异性和高效的扩增引物。 MGB缀合还增加了短寡核苷酸的鉴别能力，提供了通过短寡核苷酸增强核苷酸序列错配检测。 本文所述的MGB缀合的探针和引物有助于各种分析和诊断程序，如扩增反应，PCR，单核苷酸多态性检测，基因敲除，差异显示，荧光能量转移，水解性探针测定等。 通过允许使用具有更高特异性和更好的辨别力的较短的寡核苷酸。

    Functionalized 3-alkynyl pyrazolopyrimidine analogues as universal bases and methods of use
    18.
    发明授权
    Functionalized 3-alkynyl pyrazolopyrimidine analogues as universal bases and methods of use 有权
    官能化3-炔基吡唑并嘧啶类似物作为通用碱基和使用方法

    公开(公告)号：US09085800B2

    公开(公告)日：2015-07-21

    申请号：US13429273

    申请日：2012-03-23

    申请人： Alexei Vorobiev ,  Eugeny A. Lukhtanov ,  Noah Scarr

    发明人： Alexei Vorobiev ,  Eugeny A. Lukhtanov ,  Noah Scarr

    IPC分类号： C12Q1/68 ,  C07H21/04

    CPC分类号： C12Q1/6853 ,  C12Q1/6846 ,  C12Q2525/117

    摘要： 3-alkynyl inosine analogs and their uses as universal bases are provided. The inosine analogues can be incorporated into nucleic acid primers and probes. They do not significantly destabilize nucleic acid duplexes. As a result, the novel nucleic acid primers and probes incorporating the inosine analogues can be used in a variety of methods. The analogs function unexpectedly well as universal bases. Not only do they stabilize duplexes substantially more than hypoxanthine opposite A, C, T, and G but they are also recognized in primers by polymerases, allowing efficient amplification.

    摘要翻译： 提供3-炔基肌苷类似物及其作为通用碱基的用途。 肌苷类似物可以并入核酸引物和探针中。 它们不会使核酸双链体显着不稳定。 因此，结合肌苷类似物的新型核酸引物和探针可以以多种方法使用。 类似物作为普遍基础出人意料地起作用。 它们不仅能够稳定双链体，而且还要与A，C，T和G相反的次黄嘌呤稳定，但也可以通过聚合酶在引物中识别，从而进行有效的扩增。

    FUNCTIONALIZED 3-ALKYNYL PYRAZOLOPYRIMIDINE ANALOGUES AS UNIVERSAL BASES AND METHODS OF USE
    19.
    发明申请
    FUNCTIONALIZED 3-ALKYNYL PYRAZOLOPYRIMIDINE ANALOGUES AS UNIVERSAL BASES AND METHODS OF USE 有权
    功能性3-烷基亚氨基吡嗪类似物作为通用基础和使用方法

    公开(公告)号：US20130261014A1

    公开(公告)日：2013-10-03

    申请号：US13827456

    申请日：2013-03-14

    申请人： Alexei Vorobiev ,  Eugeny A. Lukhtanov ,  Noah Scarr

    发明人： Alexei Vorobiev ,  Eugeny A. Lukhtanov ,  Noah Scarr

    IPC分类号： C07H1/00

    CPC分类号： C07H1/00 ,  C12Q1/6816 ,  C12Q1/6846 ,  C12Q2525/117

    摘要： 3-alkynyl inosine analogs and their uses as universal bases are provided. The inosine analogues can be incorporated into nucleic acid primers and probes. They do not significantly destabilize nucleic acid duplexes. As a result, the novel nucleic acid primers and probes incorporating the inosine analogues can be used in a variety of methods. The analogs function unexpectedly well as universal bases. Not only do they stabilize duplexes substantially more than hypoxanthine opposite A, C, T, and G but they are also recognized in primers by polymerases, allowing efficient amplification.

    摘要翻译： 提供3-炔基肌苷类似物及其作为通用碱基的用途。 肌苷类似物可以并入核酸引物和探针中。 它们不会使核酸双链体显着不稳定。 因此，结合肌苷类似物的新型核酸引物和探针可以以多种方法使用。 类似物作为普遍基础出人意料地起作用。 它们不仅能够稳定双链体，而且与A，C，T和G相反的次黄嘌呤稳定，而且在引物中也被聚合酶识别，从而进行有效的扩增。

    Minor groove binder phosphoramidites and methods of use
    20.
    发明授权
    Minor groove binder phosphoramidites and methods of use 有权
    次槽粘合剂亚磷酰胺及其使用方法

    公开(公告)号：US09056887B2

    公开(公告)日：2015-06-16

    申请号：US13556513

    申请日：2012-07-24

    申请人： Alexei Vorobiev ,  Eugeny A. Lukhtanov

    发明人： Alexei Vorobiev ,  Eugeny A. Lukhtanov

    IPC分类号： C07H21/00 ,  C07F9/02 ,  C07D403/00 ,  C07F9/547 ,  C07F9/6506 ,  C07F9/6558 ,  C07F9/6561 ,  C07D235/18 ,  C07D403/04 ,  C07D519/00

    CPC分类号： C07H21/00 ,  C07D235/18 ,  C07D403/04 ,  C07D519/00 ,  C07F9/65068 ,  C07F9/65583 ,  C07F9/6561 ,  Y02P20/55

    摘要： Minor groove binder phosphoramidites having the formula M-L-PA, wherein M is a minor groove binder comprising a protected heteroaromatic amine, L is a linker, and PA is a phosphoramidite group, have been synthesized. Preferred methods of synthesis include synthesizing a minor groove binder intermediate containing a transiently protected hydroxyl group, protecting heteroaromatic amines of the corresponding minor groove binder intermediate as carbamate intermediates, reacting the carbamate intermediate to remove the transient protecting group to yield carbamate-protected minor groove binder agent as an intermediate with a free hydroxyl group, and converting the intermediate with a free hydroxyl group to the desired minor groove binder phosphoramidite. These minor groove binder phosphoramidites are useful in the preparation of oligonucleotide conjugates, particularly those for use as probes and primers. In preferred methods, an oligonucleotide sequence is synthesized using nucleoside phosphoramidites and the minor groove binder phosphoramidite is incorporated into the oligonucleotide sequence to form a protected oligonucleotide-minor groove binder conjugate. Then, deprotection produces the oligonucleotide-minor groove binder.

    摘要翻译： 具有式M-L-PA的次槽粘合剂亚磷酰胺，其中M是包含被保护的杂芳族胺的小沟粘合剂，L是接头，PA是亚磷酰胺基团。 优选的合成方法包括合成含有瞬时保护的羟基的小沟粘合剂中间体，保护相应的小沟粘合剂中间体的杂芳族胺作为氨基甲酸酯中间体，使氨基甲酸酯中间体反应以除去瞬时保护基团以产生氨基甲酸酯保护的小槽粘合剂 作为具有游离羟基的中间体，并将具有游离羟基的中间体转化为所需的小沟结合剂亚磷酰胺。 这些小沟粘结剂亚磷酰胺可用于制备寡核苷酸缀合物，特别是用作探针和引物的寡核苷酸缀合物。 在优选的方法中，使用核苷亚磷酰胺合成寡核苷酸序列，并将小沟粘合剂亚磷酰胺掺入寡核苷酸序列中以形成受保护的寡核苷酸 - 小沟粘合剂缀合物。 然后，脱保护产生寡核苷酸 - 小沟结合剂。