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1. 发明申请

US20130122506A1 NUCLEIC ACID BASE ANALOGS WITH QUENCHING AND FLUORESCENT ACTIVITIES AND APPLICATIONS THEREOF 有权
标题翻译：具有猝灭和荧光活性的核酸碱基模拟物及其应用
公开(公告)号：US20130122506A1
公开(公告)日：2013-05-16
申请号：US13642111
申请日：2011-04-21
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui , Rie Yamashige
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui , Rie Yamashige
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , C07H19/044 , C12Q1/6818 , C12Q2525/117
摘要： It is an object of the present invention to provide quenching or fluorescent nucleic acid base analogs and applications thereof. The quencher of the present invention has a 2-nitropyrrole structure represented by Formula I:[Formula 1] (in Formula I, R1 and R2 are groups independently selected from the group consisting of: ribose and deoxyribose; hydrogen, hydroxyl and SH groups, and halogens; substituted or unsubstituted alkyl, alkenyl, and alkynyl groups each having 2 to 10 carbon atoms; one or more five-membered heterocyclic rings, one or more six-membered heterocyclic rings, and one or more fused heterocyclic rings, these heterocylic rings containing nitrogen or sulfur, and one or more aromatic rings; sugars, sugar chains, amino acids, and peptides; and fluorescent molecules linked via linkers).
摘要翻译：本发明的目的是提供淬灭或荧光核酸碱基类似物及其应用。本发明的猝灭剂具有由式I表示的2-硝基吡咯结构：[式1]（式I中，R 1和R 2分别独立地选自：核糖和脱氧核糖;氢，羟基和SH基团，卤素;取代或未取代的各自具有2-10个碳原子的烷基，烯基和炔基;一个或多个五元杂环，一个或多个六元杂环和一个或多个稠合杂环，这些杂环含有氮或硫，以及一个或多个芳环;糖，糖链，氨基酸和肽;以及通过接头连接的荧光分子）。

2. 发明授权

US09285319B2 Nucleic acid base analogs with quenching and fluorescent activities and applications thereof 有权
标题翻译：具有淬灭和荧光活性的核酸碱基类似物及其应用
公开(公告)号：US09285319B2
公开(公告)日：2016-03-15
申请号：US13642111
申请日：2011-04-21
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui , Rie Yamashige
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui , Rie Yamashige
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04 , G01N21/64 , C07H19/044
CPC分类号： G01N21/6486 , C07H19/044 , C12Q1/6818 , C12Q2525/117
摘要： It is an object of the present invention to provide quenching or fluorescent nucleic acid base analogs and applications thereof. The quencher of the present invention has a 2-nitropyrrole structure represented by Formula I:[Formula 1] (in Formula I, R1 and R2 are groups independently selected from the group consisting of: ribose and deoxyribose; hydrogen, hydroxyl and SH groups, and halogens; substituted or unsubstituted alkyl, alkenyl, and alkynyl groups each having 2 to 10 carbon atoms; one or more five-membered heterocyclic rings, one or more six-membered heterocyclic rings, and one or more fused heterocyclic rings, these heterocylic rings containing nitrogen or sulfur, and one or more aromatic rings; sugars, sugar chains, amino acids, and peptides; and fluorescent molecules linked via linkers).
摘要翻译：本发明的目的是提供淬灭或荧光核酸碱基类似物及其应用。本发明的猝灭剂具有由式I表示的2-硝基吡咯结构：[式1]（式I中，R 1和R 2分别独立地选自：核糖和脱氧核糖;氢，羟基和SH基团，卤素;取代或未取代的各自具有2-10个碳原子的烷基，烯基和炔基;一个或多个五元杂环，一个或多个六元杂环和一个或多个稠合杂环，这些杂环含有氮或硫，以及一个或多个芳环;糖，糖链，氨基酸和肽;以及通过接头连接的荧光分子）。

3. 发明授权

US07745417B2 Nucleosides or nucleotides having novel unnatural bases and use thereof 有权
标题翻译：具有新的非天然碱基的核苷或核苷及其用途
公开(公告)号：US07745417B2
公开(公告)日：2010-06-29
申请号：US10521454
申请日：2003-02-28
申请人： Ichiro Hirao , Shigeyuki Yokoyama , Michiko Hirao , Tsuneo Mitsui
发明人： Ichiro Hirao , Shigeyuki Yokoyama , Michiko Hirao , Tsuneo Mitsui
IPC分类号： A61K31/70 , C07H19/00 , C07H21/00 , C07H21/02
CPC分类号： C07H11/04 , C07H7/06
摘要： The object of the present invention is to provide a nucleoside or nucleotide having an unnatural base. The nucleoside or nucleotide of the present invention has a 5-substituted-2-oxo(1H)-pyridin-3-yl group as a base. Preferably, the 5-position of the above base is substituted with a substituent selected from the group consisting of the following: 1) a photoreactive group selected from iodine and bromine; 2) an alkenyl group, an alkynyl group or an amino group, or a derivative thereof; 3) biotin or a derivative thereof; and 4) a fluorescent molecule selected from fluorescein, 6-carboxyfluorescein, tetramethyl-6-carboxyrhodamine, and derivatives thereof.
摘要翻译：本发明的目的是提供具有非天然碱基的核苷或核苷酸。本发明的核苷或核苷酸具有5-取代-2-氧代（1H） - 吡啶-3-基作为碱基。优选地，上述碱的5位被选自下列的取代基取代：1）选自碘和溴的光反应性基团; 2）烯基，炔基或氨基或其衍生物; 3）生物素或其衍生物; 和4）选自荧光素，6-羧基荧光素，四甲基-6-羧基罗丹明及其衍生物的荧光分子。

4. 发明申请

US20120245340A1 NOVEL ARTIFICIAL FLUORESCENT BASES 有权
标题翻译：新人造荧光基
公开(公告)号：US20120245340A1
公开(公告)日：2012-09-27
申请号：US13500303
申请日：2010-10-06
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui
IPC分类号： C07H19/20 , C07H21/02 , C07H19/173 , C07H21/04 , C12P19/34 , C07H19/048
CPC分类号： C07D471/04 , C07D473/32 , C07H19/173 , C07H19/23 , C09K11/06 , C09K2211/1074
摘要： The present invention relates to novel unnatural fluorescent nucleic acid bases, that is, a purine base, a 1-deazapurine base, and a 1,7-deazapurine base each having a functional group which consists of two or more heterocyclic moieties linked together, at the 6-position thereof (the 6-position of purine ring). The present invention also relates to a compound containing the unnatural base, a derivative thereof, and a nucleic acid containing a nucleotide having the unnatural base. The present invention also relates to a method of preparing the nucleic acid. The unnatural base of the present invention has excellent fluorescence characteristics and also has excellent properties as a universal base.
摘要翻译：本发明涉及新型非天然荧光核酸碱基，即嘌呤碱，1-脱氮嘌呤碱和1,7-deazapurine碱，其各自具有由两个或多个连接在一起的杂环基部分组成的官能团，在其6-位（嘌呤环的6-位）。本发明还涉及含有非天然碱，其衍生物和含有具有非天然碱基的核苷酸的核酸的化合物。本发明还涉及制备核酸的方法。本发明的非天然碱具有优异的荧光特性，也具有优异的作为通用基材的特性。

5. 发明授权

US08686130B2 Artificial fluorescent bases 有权
标题翻译：人造荧光基
公开(公告)号：US08686130B2
公开(公告)日：2014-04-01
申请号：US13500303
申请日：2010-10-06
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama , Tsuneo Mitsui
IPC分类号： C07H19/00 , C07H19/22
CPC分类号： C07D471/04 , C07D473/32 , C07H19/173 , C07H19/23 , C09K11/06 , C09K2211/1074
摘要： The present invention relates to novel unnatural fluorescent nucleic acid bases, that is, a purine base, a 1-deazapurine base, and a 1,7-deazapurine base each having a functional group which consists of two or more heterocyclic moieties linked together, at the 6-position thereof (the 6-position of purine ring). The present invention also relates to a compound containing the unnatural base, a derivative thereof, and a nucleic acid containing a nucleotide having the unnatural base. The present invention also relates to a method of preparing the nucleic acid. The unnatural base of the present invention has excellent fluorescence characteristics and also has excellent properties as a universal base.
摘要翻译：本发明涉及新型非天然荧光核酸碱基，即嘌呤碱，1-脱氮嘌呤碱和1,7-deazapurine碱，其各自具有由两个或多个连接在一起的杂环基部分组成的官能团，在其6-位（嘌呤环的6-位）。本发明还涉及含有非天然碱，其衍生物和含有具有非天然碱基的核苷酸的核酸的化合物。本发明还涉及制备核酸的方法。本发明的非天然碱具有优异的荧光特性，也具有优异的作为通用基材的特性。

6. 发明申请

US20060263771A1 Nucleoside or nucleotides having novel unnatural bases and utilization of the same 有权
标题翻译：具有新的非天然碱基的核苷或核苷酸及其利用
公开(公告)号：US20060263771A1
公开(公告)日：2006-11-23
申请号：US10521454
申请日：2003-02-28
申请人： Ichiro Hirao , Shigeyuki Yokoyama , Michiko Hirao , Tsuneo Mitsui
发明人： Ichiro Hirao , Shigeyuki Yokoyama , Michiko Hirao , Tsuneo Mitsui
IPC分类号： C12Q1/68 , C07H21/04
CPC分类号： C07H11/04 , C07H7/06
摘要： The object of the present invention is to provide a nucleoside or nucleotide having an unnatural base. The nucleoside or nucleotide of the present invention has a 5-substituted-2-oxo(1H)-pyridin-3-yl group as a base. Preferably, the 5-position of the above base is substituted with a substituent selected from the group consisting of the following: 1) a photoreactive group selected from iodine and bromine; 2) an alkenyl group, an alkynyl group or an amino group, or a derivative thereof; 3) biotin or a derivative thereof; and 4) a fluorescent molecule selected from fluorescein, 6-carboxyfluorescein, tetramethyl-6-carboxyrhodamine, and derivatives thereof.
摘要翻译：本发明的目的是提供具有非天然碱基的核苷或核苷酸。本发明的核苷或核苷酸具有5-取代-2-氧代（1H） - 吡啶-3-基作为碱基。优选地，上述碱的5位被选自下列的取代基取代：1）选自碘和溴的光反应性基团; 2）烯基，炔基或氨基或其衍生物; 3）生物素或其衍生物; 和4）选自荧光素，6-羧基荧光素，四甲基-6-羧基罗丹明及其衍生物的荧光分子。

7. 发明授权

US08895712B2 Artificial base pair capable of forming specific base pair 有权
标题翻译：能够形成特定碱基对的人造碱基对
公开(公告)号：US08895712B2
公开(公告)日：2014-11-25
申请号：US13500635
申请日：2010-10-06
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
IPC分类号： C07H21/02 , C07H21/04
CPC分类号： C07H21/02 , C07H21/04 , C12Q1/6827 , C12Q1/6832 , C12Q2527/107 , C12Q2525/117
摘要： The present invention provides a double-stranded nucleic acid in which at least one nucleic acid strand includes an unnatural base that forms a self-complementary base pair or an unnatural base that forms a base pair with any natural base with substantially the same thermal stability. The present invention also provides a method of hybridizing a first nucleic acid strand with a second nucleic acid strand, wherein the first nucleic acid strand includes an unnatural base that forms a self-complementary base pair or an unnatural base that forms a base pair with any natural base with substantially the same thermal stability, and a method of applying the nucleic acid to SNP detection, a DNA chip, DNA/RNA computing, or an in vitro translation system. The present invention provides a method of introducing an unnatural base into a nucleic acid strand and thereby controlling the thermodynamic stability in hybridization of the nucleic acid strand.
摘要翻译：本发明提供双链核酸，其中至少一个核酸链包含形成自身互补碱基对的非天然碱基或与任何具有基本上相同热稳定性的天然碱形成碱基对的非天然碱基。本发明还提供了将第一核酸链与第二核酸链杂交的方法，其中所述第一核酸链包含形成自身互补碱基对的非天然碱基或与任意形成碱基对的非天然碱基具有基本上相同的热稳定性的天然碱基，以及将核酸应用于SNP检测，DNA芯片，DNA / RNA计算或体外翻译系统的方法。本发明提供了将非天然碱引入核酸链，从而控制核酸链杂交时的热力学稳定性的方法。

8. 发明申请

US20120231462A1 ARTIFICIAL BASE PAIR CAPABLE OF FORMING SPECIFIC BASE PAIR 有权
标题翻译：人造基础配对形成特定基础对
公开(公告)号：US20120231462A1
公开(公告)日：2012-09-13
申请号：US13500635
申请日：2010-10-06
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
IPC分类号： C12Q1/68 , C07K1/00 , G01N33/566 , C07H21/02 , C07H21/04
CPC分类号： C07H21/02 , C07H21/04 , C12Q1/6827 , C12Q1/6832 , C12Q2527/107 , C12Q2525/117
摘要： The present invention provides a double-stranded nucleic acid in which at least one nucleic acid strand includes an unnatural base that forms a self-complementary base pair or an unnatural base that forms a base pair with any natural base with substantially the same thermal stability. The present invention also provides a method of hybridizing a first nucleic acid strand with a second nucleic acid strand, wherein the first nucleic acid strand includes an unnatural base that forms a self-complementary base pair or an unnatural base that forms a base pair with any natural base with substantially the same thermal stability, and a method of applying the nucleic acid to SNP detection, a DNA chip, DNA/RNA computing, or an in vitro translation system. The present invention provides a method of introducing an unnatural base into a nucleic acid strand and thereby controlling the thermodynamic stability in hybridization of the nucleic acid strand.
摘要翻译：本发明提供双链核酸，其中至少一个核酸链包含形成自身互补碱基对的非天然碱基或与任何具有基本上相同热稳定性的天然碱形成碱基对的非天然碱基。本发明还提供了将第一核酸链与第二核酸链杂交的方法，其中所述第一核酸链包含形成自身互补碱基对的非天然碱基或与任意形成碱基对的非天然碱基具有基本上相同的热稳定性的天然碱基，以及将核酸应用于SNP检测，DNA芯片，DNA / RNA计算或体外翻译系统的方法。本发明提供了将非天然碱引入核酸链，从而控制核酸链杂交时的热力学稳定性的方法。

9. 发明授权

US08212014B2 Artificial base pairs and uses thereof 有权
标题翻译：人造碱基对及其用途
公开(公告)号：US08212014B2
公开(公告)日：2012-07-03
申请号：US11989840
申请日：2006-08-04
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
IPC分类号： C07H21/00 , C07H21/02 , C12Q1/68 , C12P19/34
CPC分类号： C12Q1/6816 , C12Q1/6844 , Y10T436/143333 , C12Q2563/107 , C12Q2525/101 , C12Q2525/117
摘要： The present invention provides nucleic acids based on novel artificial base pairing, as well as a preparation method and uses thereof.In the nucleic acids of the present invention, a nucleotide having a substituted or unsubstituted 2-formyl-1H-pyrrol-1-yl group as a base forms a base pair with a nucleotide having a 6-substituted 9H-purin-9-yl group as a base. The inventive method for preparing a nucleic acid comprises effecting transcription, reverse transcription or replication by using, as a template, a nucleic acid incorporating a nucleotide having a substituted or unsubstituted 2-formyl-1H-pyrrol-1-yl group as a base, so that the nucleotide having a 6-substituted 9H-purin-9-yl group is incorporated at a site complementary to the nucleotide having a substituted or unsubstituted 2-formyl-1H-pyrrol-1-yl group.
摘要翻译：本发明提供了基于新的人造碱基配对的核酸及其制备方法和用途。在本发明的核酸中，具有取代或未取代的2-甲酰基-1H-吡咯-1-基作为碱基的核苷酸与具有6-取代的9H-嘌呤-9-基的核苷酸形成碱基对集团为基地。用于制备核酸的本发明的方法包括通过使用掺入具有取代或未取代的2-甲酰基-1H-吡咯-1-基的核苷酸作为碱的核酸作为模板来实现转录，逆转录或复制，使得具有6-取代的9H-嘌呤-9-基的核苷酸掺入与具有取代或未取代的2-甲酰基-1H-吡咯-1-基的核苷酸互补的位点。

10. 发明申请

US20140194320A1 METHOD FOR PREPARING NUCLEIC ACID APTAMER 审中-公开
标题翻译：制备核酸APTAMER的方法
公开(公告)号：US20140194320A1
公开(公告)日：2014-07-10
申请号：US14237645
申请日：2012-08-08
申请人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
发明人： Ichiro Hirao , Michiko Hirao , Shigeyuki Yokoyama
IPC分类号： C12N15/115 , C12Q1/68
CPC分类号： G01N33/543 , A61K31/7105 , C12N15/1048 , C12N15/115 , C12N2310/16 , C12N2320/13 , C12N2320/30 , C12N2330/30 , C12Q1/6834 , G01N33/5308 , G01N2333/47 , G01N2333/4703
摘要： An object of the present invention is to develop and provide a method for efficiently and conveniently producing a nucleic acid aptamer, particularly, a DNA aptamer, having high specificity for and high binding activity against a target substance. The present invention provides a method for producing a nucleic acid aptamer, comprising: a complex formation step of mixing a single-stranded nucleic acid library with a target substance in a solution to form a complex of a single-stranded nucleic acid and the target substance; an immobilization step of mixing the solution after the preceding step with a solid-phase support to immobilize the complex onto the solid-phase support via connector(s) adsorbed on the target substance and/or the solid-phase support; a recovery step of recovering the complex immobilized on the solid-phase support from the solution; an amplification step of recovering the single-stranded nucleic acid from the complex, followed by amplification by a nucleic acid amplification method; and a single-stranded nucleic acid preparation step of converting the double-stranded nucleic acids obtained in the amplification step into single strands and then forming an intramolecular conformation.
摘要翻译：本发明的目的是开发和提供一种有效且方便地制备对靶物质具有高特异性和高结合活性的核酸适体，特别是DNA适体。本发明提供了一种生产核酸适配体的方法，包括：在溶液中将单链核酸文库与目标物质混合以形成单链核酸与靶物质的复合物的复合物形成步骤 ; 固定步骤，将上述步骤之后的溶液与固相支持物混合，以通过吸附在目标物质和/或固相载体上的连接器将复合物固定在固相载体上; 从溶液中回收固定在固相载体上的络合物的回收步骤; 从复合物中回收单链核酸，然后用核酸扩增法扩增的扩增步骤; 以及将扩增步骤中得到的双链核酸转化为单链然后形成分子内构象的单链核酸制备步骤。

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