Nucleic acid mediated electron transfer

• US 6,071,699 A
• Filed: 06/19/1998
• Issued: 06/06/2000
• Est. Priority Date: 06/07/1996
• 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:
  US09/100,507
• Time in Patent Office: 718 Days
• Field of Search: 435/6, 536/24.3, 935/77, 935/78, 436/2, 436/149
• 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 2565/101 Interaction between at least two labels

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 nucleic acid sequence in a 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 nucleic acid probe covalently attached to an electrode, wherein at least one of said probe or target sequence comprises a moiety capable of donating or accepting an electron; and

b) detecting electron transfer between said moiety which donates or accepts an electron and said electrode to distinguish between single stranded and double stranded nucleic acid to detect said target nucleic acid.

2. A method according to claim 1 wherein said probe nucleic acid comprises said moiety capable of donating or accepting an electron.

3. A method according to claim 1 wherein said target nucleic acid comprises said moiety capable of donating or accepting an electron.

4. A method according to claim 1 wherein a base of said nucleic acid probe comprises said moiety capable of donating or accepting an electron.

5. A method according to claim 1 wherein said nucleic acid probe comprises a ribose-phosphate backbone.

6. A method according to claim 5 wherein a ribose of said nucleic acid probe comprises said moiety capable of donating or accepting an electron.

7. A method according to claim 5 wherein a phosphate of said nucleic acid probe comprises said moiety capable of donating or accepting an electron.

8. A method according to claim 1 wherein said nucleic acid probe comprises a nucleic acid analog.

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

10. A method according to claim 1 wherein said moiety capable of donating or accepting an electron is a transition metal complex.

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

12. A method according to claim 1 wherein an electron transport mediator is utilized in detecting said electron transfer.