Nucleic acid mediated electron transfer

• US 6,087,100 A
• Filed: 10/08/1997
• Issued: 07/11/2000
• 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:
  US08/946,679
• Time in Patent Office: 1,007 Days
• Field of Search: 536/23.1, 536/24.3, 536/24.31, 536/24.32, 536/24.33, 536/25.32, 935/5, 436/6
• 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 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 2537/101 Homogeneous assay format, e.g. one pot reaction
• 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 2565/301 Pyrophosphate (PPi)

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 single-stranded nucleic acid containing at least one electron donor moiety and at least one electron acceptor moiety, said electron donor moiety and said electron acceptor moiety being covalently attached to said nucleic acid.

2. A composition comprising a first single stranded nucleic acid comprising an electrode and a second single stranded nucleic acid comprising a covalently attached electron transfer moiety.

3. A method of detecting a target sequence in a nucleic acid sample comprising:

a) applying a first input signal to a hybridization complex comprising said target sequence, which if present, is hybridized to at least a first nucleic acid, wherein said hybridization complex has a covalently attached electron donor moiety and a 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.

4. A method according to claim 3 wherein at least one of said electron donor and said electron acceptor moieties is an electrode.

5. A method according to claim 3 wherein at least one of said electron donor and said electron acceptor moieties is a transition metal complex.

6. A method according to claim 3 wherein said electron donor moiety is an electrode and said electron acceptor moiety is a transition metal complex.

7. A method according to claim 3 wherein said first nucleic acid comprises a covalently attached electron donor moiety and a covalently attached electron acceptor moiety.

8. A method according to claim 3 wherein said first nucleic acid comprises a covalently attached electron donor moiety and said target sequence comprises a covalently attached electron acceptor moiety.

9. A method according to claim 3 wherein at least one of said electron donor and said electron acceptor moieties is attached to a terminal base.

10. A method according to claim 3 wherein said nucleic acid comprises a ribose-phosphate backbone.

11. A method according to claim 10 wherein at least one of said electron donor and said electron acceptor moieties is attached to a ribose of said ribose-phosphate backbone.

12. A method according to claim 10 wherein at least one of said electron donor and said electron acceptor moieties is attached to a phosphate of said ribose-phosphate backbone.

13. A method according to claim 3 wherein said nucleic acid is a nucleic acid analog.

14. A method according to claim 13 wherein said nucleic acid analog is peptide nucleic acid.

15. A method according to claim 14 wherein said transition metal complex comprises ruthenium, rhenium, osmium, platinum, copper or iron.

16. A method according to claim 3 wherein at least one of said electron donor and said electron acceptor moieties is an organic electron donor or acceptor.