Nucleic acid mediated electron transfer

• US 5,824,473 A
• Filed: 06/07/1995
• Issued: 10/20/1998
• 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): Jones, W. Gary
• Assistant Examiner(s): REES, DIANE
• Application Number:
  US08/475,051
• Time in Patent Office: 1,231 Days
• Field of Search: 435/6, 435/5, 435/91.2, 536/24.3, 536/26.6, 536/23.1, 536/24.33, 536/24.32
• 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 method of detecting a target sequence in a nucleic acid sample comprising:

a) initiating electron transfer by applying an initiation signal to a hybridization complex comprising said target sequence, if present, hybridized to a single-stranded probe nucleic acid containing at least one electron donor moiety and at least one electron acceptor moiety, wherein said electron donor moiety and said electron acceptor moiety are covalently attached to said probe nucleic acid, and wherein said electron donor moiety and said electron acceptor moiety are not redox proteins; and

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

5. A method according to claim 1, 2, 3 or 4 wherein at least one of said electron acceptor moiety and said electron donor moiety is attached to a terminal base.

6. A method according to claim 1, 2, 3 or 4 wherein said nucleic acid comprises a ribose-phosphate backbone.

7. A method according to claim 6 wherein at least one of said electron acceptor moiety and said electron donor moiety is attached to a ribose of said ribose-phosphate backbone.

8. A method according to claim 1, 2, 3 or 4 wherein said nucleic acid comprises a nucleic acid analog.

9. A composition according to claim 8 wherein said nucleic acid analog comprises a peptide nucleic acid.

10. A method according to claim 1, 2, 3 or 4 wherein at least one of said electron donor moiety and said electron acceptor moiety is a transition metal complex.

11. A composition according to claim 10 wherein said transition metal complex comprises ruthenium, rhenium, osmium, copper or iron.

12. A method according to claim 1, 2, 3 or 4 wherein at least one of said electron donor moiety and said electron acceptor moiety is an organic electron donor or acceptor.

13. A method according to claim 1 or 3 wherein at least one of said electron donor moiety and said electron acceptor moiety is an electrode.

14. A method according to claim 1, 2, 3 or 4 wherein said detection comprises electronic detection.

15. A method according to claim 1, 2, 3 or 4 wherein said detection comprises fluorescence detection.

16. A method according to claim 1, 2, 3 or 4 wherein said detection comprises absorbance detection.

17. A method according to claim 1, 2, 3 or 4 wherein said initiation signal comprises applied potential.

18. A method according to claim 1, 2, 3 or 4 wherein said initiation signal comprises photoactivation.

19. A method according to claim 1, 2, 3 or 4 wherein said initiation signal comprises applied potential and photoactivation.

20. A method according to claim 1, 2, 3 or 4 wherein said initiation signal comprises applied potential and said detection comprises electronic detection.

21. A method according to claim 1 or 2 further comprising the step of forming the hybridization complex by hybridizing said probe nucleic acid to said target nucleic acid.

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

a) initiating electron transfer by applying an initiation signal to a hybridization complex comprising said target sequence, if present, hybridized to a single-stranded probe nucleic acid containing 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 an electron transfer moiety which is not a redox protein and which is covalently attached to said nucleic acid; and

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

3. A method of detecting a target sequence in a nucleic acid wherein said target sequence comprises a first target domain and a second target domain adjacent to said first target domain, wherein said method comprises:

a) initiating electron transfer by applying an initiation signal to a hybridization complex comprising a first probe nucleic acid containing at least one covalently attached electron donor moiety hybridized to said first target domain and a second probe nucleic acid containing at least one covalently attached electron acceptor moiety hybridized to said second target domain, wherein said electron donor and acceptor moieties are not redox proteins; and

b) detecting electron transfer between said electron donor and said electron acceptor moieties.

22. A method according to claim 3 or 4 further comprising the step of forming the hybridization complex by hybridizing said first probe nucleic acid to said first target domain and hybridizing said second probe nucleic acid to said second target domain.

4. A method of detecting a target sequence in a nucleic acid wherein said target sequence comprises a first target domain and a second target domain adjacent to said first target domain, wherein said method comprises:

a) initiating electron transfer by applying an initiation signal to a hybridization complex comprising a first probe nucleic acid containing at least one electron donor moiety hybridized to said first target domain and a second probe nucleic acid containing at least one electron acceptor moiety hybridized to said second target domain, wherein at least one of said electron donor moiety and said electron acceptor moiety is an electrode, and the other is an electron transfer moiety which is not a redox protein and which is covalently attached to said nucleic acid; and

b) detecting electron transfer between said electron donor and said electron acceptor moieties.