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RNA-selective hybridization reagent and use of the same

外国特許コード F110005402
整理番号 K02703WO
掲載日 2011年9月5日
出願国 アメリカ合衆国
出願番号 92190909
公報番号 20110033863
公報番号 8609826
出願日 平成21年3月11日(2009.3.11)
公報発行日 平成23年2月10日(2011.2.10)
公報発行日 平成25年12月17日(2013.12.17)
国際出願番号 JP2009054675
国際公開番号 WO2009113580
国際出願日 平成21年3月11日(2009.3.11)
国際公開日 平成21年9月17日(2009.9.17)
優先権データ
  • 特願2008-061751 (2008.3.11) JP
  • 2009WO-JP54675 (2009.3.11) WO
発明の名称 (英語) RNA-selective hybridization reagent and use of the same
発明の概要(英語) (US8609826)
Provided is a nucleoside derivative which has a high affinity for RNA.
Use is made of a nucleoside derivative represented by either formula (1) or formula (2). (In formulae (1) and (2), Z represents a carbon atom or a nitrogen atom; R1 represents a hydrogen atom or a hydroxyl-protecting group; and R2 represents a hydrogen atom or a phosphodiester group).
特許請求の範囲(英語) [claim1]
1. An RNA hybridization reagent provided with one species or two or more species of nucleotide derivative units represented by any of the following Formulae (5) and (6):
where Z represents a carbon atom or a nitrogen atom; X1 represents 0, S or Se; X2 represents SH (or S-), S or Se-, or an alkyl group having 1 to 4 carbons or a morpholino group,
wherein the RNA hybridization reagent selectively hybridizes RNA having a base sequence containing U corresponding to the Formula (5) and/or A corresponding to the Formula (6).
[claim2]
2. The RNA hybridization reagent according to claim 1, wherein the nucleotide derivative unit is represented by the Formula (5) and the Z is a nitrogen atom.
[claim3]
3. The RNA hybridization reagent according to claim 2, provided with the nucleotide derivative unit at an extremity.
[claim4]
4. The RNA hybridization reagent according to claim 1, having a base sequence which forms a stem-loop structure, and provided with the nucleotide derivative unit in the loop.
[claim5]
5. A probe set for detecting a mutation on an RNA, comprising: a first probe provided with one species or two or more species of nucleotide derivative units represented by any of the following Formulae (5) and (6) at the 5' end or the 3' end corresponding to a site of the mutation; and
one species or two or more species of second probes provided with a deoxynucleotide having a base complementary to a base that has the possibility to be present in a site of the mutation at the 3' end or the 5' end corresponding to the site of the mutation:
where Z represents a carbon atom or a nitrogen atom; X1 represents 0, S or Se; X2 represents SH (or S-), S or Se-, or an alkyl group having 1 to 4 carbons or a morpholino group,
wherein the first probe selectively hybridizes RNA having a base sequence containing U corresponding to the Formula (5) and/or A corresponding to the Formula (6).
[claim6]
6. A detecting method for a single base polymorphism, comprising: preparing an RNA sample as a gene expression product having the possibility of containing the single base polymorphism;
with a probe set being provided containing a first probe provided with a nucleotide derivative unit represented by any of the following Formulae (5) and (6) at the 5' end or the 3' end corresponding to a site of the single base polymorphism, and one species or two or more species of second probes provided with a deoxynucleotide having a base complementary to a base that has the possibility to be present in a site of the single base polymorphism at the 3' end or the 5' end corresponding to the site of the single base polymorphism,
causing the first probe, the second probe and the RNA sample to contact one another allowing hybridization in combinations of one species or two or more species obtained by combining one species of the first probe and one species of the second probe selected from the probe set,
where Z represents a carbon atom or a nitrogen atom; X' represents 0, S or Se; X2 represents SH (or S-), S or Se-, or an alkyl group having 1 to 4 carbons or a morpholino group; and
detecting a fluorescence signal based on the first probe which is a hybridization product among the RNA sample, the first probe and the second probe,
wherein the first probe selectively hybridizes the RNA sample having a base sequence containing U corresponding to the Formula 5 and/or A corresponding to the Formula (6).
[claim7]
7. The RNA hybridization reagent according to claim 2, provided with the nucleotide derivative unit at an extremity.
[claim8]
8. The RNA hybridization reagent according to claim 2, having a base sequence which forms a stem-loop structure, and provided with the nucleotide derivative unit in the loop.
[claim9]
9. The RNA hybridization reagent according to claim 3, having a base sequence which forms a stem-loop structure, and provided with the nucleotide derivative unit in the loop.
[claim10]
10. The RNA hybridization reagent according to claim 7, having a base sequence which forms a stern-loop structure, and provided with the nucleotide derivative unit in the loop.
[claim11]
11. The RNA hybridization reagent according to claim 1, wherein the Z is a nitrogen atom.
[claim12]
12. The RNA hybridization reagent according to claim 1, wherein the RNA hybridization reagent has one or more deoxyribonucleotides with natural bases as one or more units other than the one species or two or more species of nucleotide derivative units represented by any of the formulae (5) and (6).
[claim13]
13. A method for forming a hybridized product with RNA comprising: contacting the RNA hybridization reagent according to claim 1 with an RNA which has a base sequence containing U corresponding to the Formula (5) and/or A corresponding to the Formula (6).
[claim14]
14. The method according to claim 13, wherein the RNA hybridization reagent has one or more deoxyribonucleotides with natural bases as one or more units other than the one species or two or more species of nucleotide derivative units represented by any of the Formulae (5) and (6).
  • 発明者/出願人(英語)
  • UENO YOSHIHITO
  • KITADE YUKIO
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
米国特許分類/主・副
  • C07D487/14
  • C07H019/173
  • C12Q001/68B8
参考情報 (研究プロジェクト等) PRESTO RNA and biofunctions AREA
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