Nucleic acid mediated electron transfer

• US 6,177,250 B1
• Filed: 05/07/1999
• Issued: 01/23/2001
• Est. Priority Date: 12/10/1993
• Status: Expired due to Term

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Fraser, Scott E., Kayyem, Jon Faiz, Meade, Thomas J.
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US09/306,737
• Time in Patent Office: 627 Days
• Field of Search: 435/6, 536/24.3, 536/23.1
• US Class Current: 435/6.11
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 23/00 Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12 esters with inorganic acids C07H11/00; metal salts, see parent compounds
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2563/113 the label being electroactive, e.g. redox labels
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label
• CPC Class Codes: C12Q 2565/101 Interaction between at least two labels
• CPC Class Codes: C12Q 2565/1015 labels being on the same oligonucleotide
• CPC Class Codes: C12Q 2565/401 Signal amplification by chemical polymerisation
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode

Abstract

The present invention provides for the selective covalent modification of nucleic acids with redox active moieties such as transition metal complexes. Electron donor and electron acceptor moieties are covalently bound to the ribose-phosphate backbone of a nucleic acid at predetermined positions. The resulting complexes represent a series of new derivatives that are bimolecular templates capable of transferring electrons over very large distances at extremely fast rates. These complexes possess unique structural features which enable the use of an entirely new class of bioconductors and photoactive probes.

Claims

1. A composition comprising:

a) a single stranded nucleic acid comprising at least one covalently attached electron donor moiety and at least one covalently attached electron acceptor moiety; and

b) an electron transfer mediator.

5. A composition according to claim 1, 2, 3 or 4 wherein said electron transport mediator is free in solution.

6. A composition according to claim 1, 2, 3 or 4 wherein said electron transport mediator is embedded in a gel.

2. A composition comprising:

a) a single stranded nucleic acid comprising at least one electron donor moiety and at least one electron acceptor moiety, wherein at least one of said electron donor moiety and said electron acceptor moiety is an electrode and the other is a covalently attached electron transfer moiety; and

b) an electron transfer mediator.

3. A composition comprising:

a) a first nucleic acid comprising at least one electron donor moiety;

b) a second nucleic acid containing at least one electron acceptor moiety;

wherein at least one of said electron donor moiety and said electron acceptor moiety is an electrode and the other is a covalently attached electron transfer moiety; and

c) an electron transfer mediator.

4. A composition comprising:

a) a first nucleic acid comprising at least one covalently attached electron donor moiety;

b) a second nucleic acid containing at least one covalently attached electron acceptor moiety; and

c) an electron transfer mediator.

7. A method for detecting the presence of a sequence in a nucleic acid sample, said method comprising:

a) applying an input signal to said sample, wherein said sample comprises;

i) an electron transfer mediator; and

ii) a hybridization complex comprising;

1) a first single stranded nucleic acid;

2) a target sequence;

wherein said hybridization complex comprises at least one covalently attached electron donor moiety and at least one covalently attached electron acceptor moiety; and

b) detecting electron transfer between said electron donor moiety and said electron acceptor moiety as an indication of the presence or absence of said target sequence.

8. A method according to claim 7 wherein at least one of said electron acceptor and electron donor moieties is an electrode.

9. A method according to claim 8 wherein at least one of said electron acceptor and electron donor moieties is a transition metal complex.

10. A method according to claim 9 wherein said transition metal complex comprises a metal selected from the group consisting of ruthenium, rhenium, osmium, platinum, copper and iron.

11. A method according to claim 8 wherein at least one of said electron acceptor and electron donor moieties is an organic electron transfer moiety.

12. A method according to claim 7 wherein said first nucleic acid comprises a covalently attached electron donor moiety and a covalently attached electron acceptor moiety.

13. A method according to claim 7 wherein said first nucleic acid comprises a covalently attached electron donor moiety and said target sequence comprises a covalently attached electron acceptor moiety.

14. A method according to claim 7 wherein said input signal comprises an AC input signal.

15. A method according to claim 14 wherein said AC input signal is applied at a plurality of frequencies.

16. A method according to claim 14 wherein said detecting is done by detecting an output signal characteristic of electron transfer between said moiety and said electrode.

17. A method according to claim 16 wherein said output signal is a current.

18. A method for detecting the presence of a sequence in a nucleic acid sample, said method comprising:

a) applying an AC input signal to said sample, wherein said sample comprises;

i) an electron transfer mediator; and

ii) a hybridization complex comprising;

1) a first single stranded nucleic acid;

2) a target sequence;

wherein said hybridization complex comprises at least one covalently attached electron transfer moiety and an electrode; and

b) detecting electron transfer between said electron transfer moiety and said electrode as an indication of the presence or absence of said target sequence.

19. A method for detecting the presence of a nucleic acid with a covalently attached electron transfer moiety in a sample, said method comprising:

a) applying an AC input signal to said sample, wherein said sample comprises;

i) an electron transfer mediator;

ii) an electrode; and

ii) said nucleic acid with a covalently attached electron transfer moiety;

b) detecting electron transfer between said electron donor moiety and said electrode.