Nucleic acid mediated electron transfer

• US 7,851,146 B2
• Filed: 10/23/2002
• Issued: 12/14/2010
• Est. Priority Date: 12/10/1993
• Status: Expired due to Fees

• 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): MUMMERT, STEPHANIE KANE
• Application Number:
  US10/279,742
• Publication Number:
  US 20030170677A1
• Time in Patent Office: 2,974 Days
• Field of Search: 435/6, 435/91.2, 536/5, 536/24.3, 536/26.6, 536/23.1
• 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 sequence, comprising

a) hybridizing a target nucleic acid comprising a first and second domain to a first single stranded nucleic acid covalently attached to a first electron transfer moiety comprising an electrode, wherein said first single stranded nucleic acid hybridizes to said first domain;

b) hybridizing a second single stranded nucleic acid to said second domain to form a hybridization complex comprising said target nucleic acid, said first single stranded nucleic acid, and said second single stranded nucleic acid, wherein said hybridization complex further comprises a second electron transfer moiety; and

c) detecting electron transfer between said second electron transfer moiety and said electrode as an indicator of the presence of said target nucleic acid in said hybridization complex.

2. The method of claim 1, wherein said second electron transfer moiety is covalently attached to said second single stranded nucleic acid.

3. The method of claim 2, wherein said second single stranded nucleic acid comprises multiple electron transfer moieties.

4. The method of claim 2 or 3, wherein said electron transfer moiety comprises a transition metal complex.

5. The method of claim 4, wherein said transition metal is selected from the group consisting of Cd, Mg, Cu, Co, Pd, Zn, Fe, and Ru.

6. The method of claim 4, wherein said transition metal complex is covalently attached to a ribose of the ribose phosphate backbone of said second single stranded nucleic acid.

7. The method of claim 6, wherein said attachment is at the 2′

or 3′

position of said ribose.

8. The method of claim 7, wherein said attachment is at the 2′

position of said ribose.