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RNA interference induction element and use thereof UPDATE

外国特許コード F110005369
整理番号 K01804WO
掲載日 2011年9月5日
出願国 アメリカ合衆国
出願番号 92050806
公報番号 20090215644
公報番号 7847089
出願日 平成18年5月15日(2006.5.15)
公報発行日 平成21年8月27日(2009.8.27)
公報発行日 平成22年12月7日(2010.12.7)
国際出願番号 JP2006310079
国際公開番号 WO2006123800
国際出願日 平成18年5月15日(2006.5.15)
国際公開日 平成18年11月23日(2006.11.23)
優先権データ
  • 特願2005-145876 (2005.5.18) JP
  • 2006WO-JP310079 (2006.5.15) WO
発明の名称 (英語) RNA interference induction element and use thereof UPDATE
発明の概要(英語) (US7847089)
The present invention provides an RNA interference induction element containing a nucleotide sequence selected from among the nucleotide sequences (a) to (c) below: (a) a nucleotide sequence containing SEQ ID NO:1 or a sequence complementary thereto; (b) a nucleotide sequence containing at least 15 continuous nucleotides present in the nucleotide sequence (a) above, and possessing RNA interference induction potential; (c) a nucleotide sequence having a homology of at least 70% to any one of the nucleotide sequences (a) and (b) above, and possessing RNA interference induction potential.
Using the RNA interference induction element of the present invention, it is easily possible to knock down a desired target gene, and to produce a siRNA for a desired target gene.
特許請求の範囲(英語) [claim1]
1. An isolated RNA interference induction element consisting of the nucleotide sequence of SEQ ID NO:1 or a sequence complementary to the full length nucleotide sequence of SEQ ID NO: 1 and with the identical length.
[claim2]
2. An isolated polynucleotide comprising i) the element of claim 1, and
ii) a nucleotide sequence comprising at least 15 continuous nucleotides present in the nucleotide sequence that encodes the transcript of a target gene, or a sequence complementary thereto, wherein the nucleotide sequence of step ii) is connected to the element so that RNA interference induction potential for the target gene can be exhibited.
[claim3]
3. The isolated polynucleotide of claim 2, wherein the nucleotide sequence of step ii) is connected to the 5' side of the element.
[claim4]
4. The isolated polynucleotide of claim 2, which comprises plural copies of the element as connected in tandem.
[claim5]
5. A vector harboring plural copies of the element of claim 1 connected in tandem.
[claim6]
6. A vector harboring i) the element of claim 1, and
ii) a promoter joined to the element so that the expression of the element can be controlled.
[claim7]
7. A vector harboring i) the element of claim 1, and
ii) at least one cloning site connected to the element so that RNA interference induction potential for a target gene can be exhibited when a nucleotide sequence comprising at least 15 continuous nucleotides present in the nucleotide sequence that encodes the transcript of the target gene or a sequence complementary thereto is inserted to the cloning site.
[claim8]
8. The vector of claim 7, wherein the cloning site is connected to the 5' side of the element.
[claim9]
9. The vector of claim 7, which further harbors a promoter joined to the element or the cloning site so that the expression of the element and the cloning site can be controlled.
[claim10]
10. A vector harboring the polynucleotide of claim 2.
[claim11]
11. The vector of claim 10, which further harbors a promoter joined to the polynucleotide so that the expression of the polynucleotide can be controlled.
[claim12]
12. An isolated cell incorporating the polynucleotide of claim 2.
[claim13]
13. An isolated cell incorporating the vector of claim 5.
[claim14]
14. A method of producing a cell wherein the expression of a target gene is suppressed, which comprises a step for transferring the polynucleotide of claim 2 into cells, and a step for selecting a cell incorporating the polynucleotide.
[claim15]
15. A method of suppressing the expression of a target gene, which comprises a step for transferring the polynucleotide of claim 2 into cells.
[claim16]
16. A method of producing a siRNA for a target gene, which comprises a step for transferring the polynucleotide of claim 2 into cells, and a step for obtaining the siRNA for the target gene from the cells incorporating the polynucleotide or the vector.
[claim17]
17. An RNA interference inducing agent comprising the polynucleotide of claim 2.
[claim18]
18. A gene knockdown polynucleotide library comprising a plurality of polynucleotides, each of which comprises a nucleotide sequence comprising at least 15 continuous nucleotides present in the nucleotide sequence that encodes each of the transcripts of a plurality of genes or a sequence complementary thereto, wherein each nucleotide sequence is connected to the element of claim 1 so that RNA interference induction potential for the gene can be exhibited.
[claim19]
19. The library of claim 18, wherein the each polynucleotide is harbored in a vector.
[claim20]
20. A cellular population incorporating the library of claim 18.
[claim21]
21. A method of screening for a functional gene, which comprises the steps (a) to (c) below: (a) analyzing the phenotype of a cellular population incorporating the library of claim 18;
(b) isolating cells with an altered phenotype from the cellular population; and
(c) obtaining a functional gene based on a nucleotide sequence in the polynucleotide or the vector incorporated in the isolated cells.
[claim22]
22. An isolated RNA-dependent RNA synthesis reaction induction element consisting of the nucleotide sequence of SEQ ID NO:1 or a sequence complementary to the full length nucleotide sequence of SEQ ID NO:1.
[claim23]
23. A method of synthesizing an RNA, which comprises the steps shown below: (a) a step for providing a template for an RNA-dependent RNA synthesis reaction comprising the element of claim 22;
(b) a step for bringing the template of (a) into contact with RNA-dependent RNA polymerase to cause the RNA-dependent RNA synthesis reaction.
[claim24]
24. An isolated gene expression suppression element consisting of the below nucleotide sequence of SEQ ID NO:1 or a sequence complementary to the full length nucleotide sequence of SEQ ID NO:1.
[claim25]
25. A method of producing a cell wherein the expression of a target gene is suppressed, which comprises a step for transferring the vector of claim 10 into cells, and a step for selecting a cell incorporating the polynucleotide or the vector.
[claim26]
26. A method of suppressing the expression of a target gene, which comprises a step for transferring the vector of claim 10 into cells.
[claim27]
27. A method of producing a siRNA for a target gene, which comprises a step for transferring the vector of claim 10 into cells, and a step for obtaining the siRNA for the target gene from the cells incorporating the polynucleotide or the vector.
[claim28]
28. An RNA interference inducing agent comprising the vector of claim 10.
[claim29]
29. An isolated cell incorporating the vector of claim 7.
[claim30]
30. A method of producing a cell wherein the expression of a target gene is suppressed, which comprises a step of transferring the vector of claim 7 into cells, and a step for selecting a cell incorporating the vector.
[claim31]
31. A method for producing siRNA for a target gene, which comprises a step for transferring the vector of claim 7 into cells, and a step for obtaining the siRNA for the target gene from the cells incorporating the vector.
[claim32]
32. A method of screening for a functional gene, which comprises the steps (a) to (c) below: (a) analyzing the phenotype of a cellular population incorporating the library of claim 20;
(b) isolating cells with an altered phenotype from the cellular population; and
(c) obtaining a functional gene based on a nucleotide sequence in the vector incorporated in the isolated cells.
  • 発明者/出願人(英語)
  • ISHII KOJIRO
  • TAKAHASHI KOHTA
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • KURUME UNIVERSITY
国際特許分類(IPC)
米国特許分類/主・副
  • C12N015/11
  • C12N015/11M
  • M12N310/14
  • M12N320/50
  • M12N330/10
  • M12N330/30
参考情報 (研究プロジェクト等) PRESTO Information and Cell Function AREA
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