Method for evaluating redox activity of nucleic acid molecule and nucleic acid molecule having redox activity

Patent metadata

Field	Value
Publication number	US-9637737-B2
Application number	US-201214129392-A
Country	US
Kind code	B2
Filing date	Jul 2, 2012
Priority date	Jul 4, 2011
Publication date	May 2, 2017
Grant date	May 2, 2017

Abstract

The present invention provides a novel technique by which the redox activity of a nucleic acid molecule can be evaluated. An evaluation method of the present invention includes: a detection step of electrochemically detecting a redox reaction to a substrate, the redox reaction being catalyzed by a nucleic acid molecule to be evaluated, using a device that electrochemically detects a redox reaction; and an evaluation step of evaluating redox activity of the nucleic acid molecule from a result of the detection of the redox reaction. As the device, a device in which a base provided with a detection portion is included, the detection portion includes an electrode system, and the nucleic acid molecule to be evaluated is arranged on the base is used. In the present invention, it is preferred that a plurality of kinds of nucleic acid molecule to be evaluated is arranged on the base, and the plurality of kinds of nucleic acid molecules to be evaluated is evaluated by a single device.

First claim

The invention claimed is:

1. A method for evaluating redox activity of a nucleic acid molecule, the method comprising:
a detection step of electrochemically detecting a redox reaction to a substrate using a device that electrochemically detects a redox reaction, wherein the redox reaction is catalyzed by at least one nucleic acid molecule to be evaluated; and
an evaluation step of evaluating redox activity of the at least one nucleic acid molecule from a result of the detection of the redox reaction, wherein
the device includes a base provided with a detection portion,
the detection portion includes an electrode system, and
the nucleic acid molecule to be evaluated is arranged, via a linker of polynucleotide having a length of from 20 to 30 bases, on the base.

2. The method according to claim 1 , wherein the at least one nucleic acid molecule to be evaluated comprises a plurality of kinds of nucleic acid molecules.

3. The method according to claim 2 , wherein the base is a microarray in which the plurality of kinds of nucleic acid molecules is arranged.

4. The method according to any one of claims 1 to 3 , wherein the detection step is a step of detecting a redox reaction catalyzed by the nucleic acid molecule in the presence of the substrate.

5. The method according to any one of claims 1 to 3 , wherein a nucleic acid aptamer capable of binding to a target is bound to the nucleic acid molecule to be evaluated.

6. The method according to claim 5 , wherein the detection step is a step of detecting a redox reaction catalyzed by the nucleic acid molecule in the presence of the substrate and the target.

7. The method according to claim 5 , wherein the detection step comprises:
a step of detecting the redox reaction in the presence of the substrate and in the absence of the target; and
a step of detecting the redox reaction in the presence of the substrate and the target, and
in the evaluation step evaluating the redox activity, in the presence of the target, of a nucleic acid molecule that does not exert the redox activity in the absence of the target.

8. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to any one of claims 1 to 3 using a device; and
screening a nucleic acid molecule having redox activity.

9. The method according to claim 4 , wherein a nucleic acid aptamer capable of binding to a target is bound to the nucleic acid molecule to be evaluated.

10. The method according to claim 9 , wherein the detection step is a step of detecting a redox reaction catalyzed by the nucleic acid molecule in the presence of the substrate and the target.

11. The method according to claim 9 , wherein
the detection step comprises:
a step of detecting the redox reaction in the presence of the substrate and in the absence of the target; and
a step of detecting the redox reaction in the presence of the substrate and the target, and
in the evaluation step, the redox activity in the presence of the target, of a nucleic acid molecule that does not exert the redox activity in the absence of the target is evaluated.

12. The method according to claim 6 , wherein the detection step comprises:
a step of detecting the redox reaction in the presence of the substrate and in the absence of the target; and
a step of detecting the redox reaction in the presence of the substrate and the target, and
in the evaluation step, the redox activity in the presence of the target, of a nucleic acid molecule that does not exert the redox activity in the absence of the target is evaluated.

13. The method according to claim 10 , wherein the detection step comprises:
a step of detecting the redox reaction in the presence of the substrate and in the absence of the target; and
a step of detecting the redox reaction in the presence of the substrate and the target, and
in the evaluation step, the redox activity in the presence of the target, of a nucleic acid molecule that does not exert the redox activity in the absence of the target is evaluated.

14. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 4 using a device; and
screening a nucleic acid molecule having redox activity.

15. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 5 using a device; and
screening a nucleic acid molecule having redox activity.

16. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 9 using a device; and
screening a nucleic acid molecule having redox activity.

17. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 6 using a device; and
screening a nucleic acid molecule having redox activity.

18. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 10 using a device; and
screening a nucleic acid molecule having redox activity.

19. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 7 using a device; and
screening a nucleic acid molecule having redox activity.

20. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 11 using a device; and
screening a nucleic acid molecule having redox activity.

21. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 12 using a device; and
screening a nucleic acid molecule having redox activity.

22. A method for screening a nucleic acid molecule having redox activity, the method comprising:
evaluating redox activity of at least one nucleic acid molecule to be evaluated by the method according to claim 13 using a device; and
screening a nucleic acid molecule having redox activity.

Assignees

• Kaneko Naoto
• Horii Katsunori
• Akitomi Jou
• Katou Shintarou
• Waga Iwao
• Nec Solution Innovators Ltd

Inventors

• Kaneko Naoto
• Horii Katsunori
• Akitomi Jou
• Katou Shintarou
• Waga Iwao

Classifications

• C12Q1/6825

  Nucleic acid detection involving sensors · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• C12N15/115

  Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers} · CPC title

• C12N2310/16

  Aptamers · CPC title

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title