Nucleic acid mediated electron transfer

• US 6,180,352 B1
• Filed: 12/10/1999
• Issued: 01/30/2001
• Est. Priority Date: 12/10/1993
• Status: Expired due to Term

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Kayyem, Jon Faiz, Meade, Thomas J., Fraser, Scott E.
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US09/459,191
• Time in Patent Office: 417 Days
• Field of Search: 435/6, 536/23.1, 536/24.3
• US Class Current: 435/6.11
• 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
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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 method of detecting the presence of a sequence in a nucleic acid sample, said method comprising:

a) applying an input signal to an electrode comprising a hybridization complex, wherein said hybridization complex comprises;

1) a single stranded nucleic acid;

2) a target sequence hybridized to said single stranded nucleic acid; and

3) at least one covalently attached electron donor moiety;

such that an electron is transferred from said electron donor moiety to said electrode such that the redox state of said electron donor moiety is changed to form an electron acceptor moiety;

b) transferring an electron from a reductant to said electron acceptor moiety; and

c) detecting electron transfer between said electron donor moiety and said electrode, wherein the presence of electron transfer is an indication of the presence of said target sequence.

2. A method according to claim 1 wherein said detecting is done by detecting an output signal characteristic of electron transfer between said donor moiety and said electrode.

5. A method according to claim 2 or 4 wherein said output signal is a current.

6. A method according to claim 1 or 3 wherein said single stranded nucleic acid comprises said electrode.

7. A method according to claim 1 or 3 wherein said single stranded nucleic acid comprises said electron donor moiety.

8. A method according to claim 1 or 3 wherein said target sequence comprises said electron donor moiety.

9. A method according to claim 1 or 3 wherein said hybridization complex comprises a plurality of electron donor moieties.

10. A method according to claim 1 or 3 wherein said electron donor moiety is a transition metal complex.

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

12. A method according to claim 1 or 3 wherein said electron donor moiety is an organic electron donor moiety.

13. A method according to claim 1 wherein said reductant is a hexacyano iron complex.

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

a) applying an input signal to an electrode comprising a hybridization complex, wherein said hybridization complex comprises;

1) a single stranded nucleic acid;

2) a target sequence hybridized to said single stranded nucleic acid; and

3) at least one covalently attached electron acceptor moiety;

such that an electron is transferred from said electron acceptor moiety to said electrode such that the redox state of said electron acceptor moiety is changed to form an electron donor moiety;

b) transferring an electron from an oxidant to said electron donor moiety; and

c) detecting electron transfer between said electron acceptor moiety and said electrode, wherein the presence of electron transfer is an indication of the presence of said target sequence.

4. A method according to claim 3 wherein said detecting is done by detecting an output signal characteristic of electron transfer between said acceptor moiety and said electrode.