Nucleosides comprising polydentate ligands

• US 6,444,423 B1
• Filed: 11/13/1998
• Issued: 09/03/2002
• Est. Priority Date: 06/07/1996
• Status: Expired due to Fees

• Current Assignee: GE Healthcare SV Corp (Thermo Fisher Scientific Incorporated)
• Original Assignee: Molecular Dynamics Inc. (General Electric Company)
• Inventors: Meade, Thomas J., Welch, Thomas W.
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US09/191,785
• Time in Patent Office: 1,390 Days
• Field of Search: 435/6, 536/22.1, 536/23.1, 536/25.3
• US Class Current: 435/6.11
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• 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: C12Q 1/6825 Nucleic acid detection involving sensors

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 nucleoside containing a covalently attached polydentate ligand, said ligand attached at the 2′

or 3′

position of said nucleoside.

2. A nucleoside according to claim 1 wherein said attachment is at the 2′

position.

3. A nucleoside according to claim 1 wherein said attachment is at the 3′

position.

5. A composition according to claim 1 further comprising a transition metal chelated to said polydentate nucleoside.

4. A phosphoramidite nucleoside containing a covalently attached polydentate ligand, said ligand attached at the 2′

position of said nucleoside.

6. A composition comprising a single-stranded 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 nucleic acid, and wherein at least one of said electron donor and said electron acceptor moieties is attached to a polydentate nucleoside.

8. A composition according to claim 6 or 7, wherein both of said electron donor and acceptor moieties are attached via polydentate nucleoside.

9. A composition according to claim 6 or 7, further comprising a transition metal chelated to said polydentate nucleoside.

10. A composition according to claim 6 or 7, wherein the other of said electron donor and acceptor moieties is an electrode.

11. A composition according to claim 6 or 7 wherein at least one of said electron donor moiety and said electron acceptor moiety is an organic electron donor or acceptor.

7. A composition comprising a first single stranded nucleic acid containing at least one covalently attached electron donor moiety and a second single stranded nucleic acid containing at least one covalently attached electron acceptor moiety, wherein at least one of said electron donor and acceptor moieties is attached to a polydentate nucleoside.

12. 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 one single stranded nucleic acid, wherein said hybridization complex has a covalently attached electron donor moiety and a covalently attached electron acceptor moiety, wherein at least one of said electron donor and acceptor moieties is attached to a polydentate nucleoside; and

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

13. A method according to claim 12 wherein said single stranded nucleic acid comprises said electron donor moiety and said electron acceptor moiety.

14. A method according to claim 12 wherein said target sequence comprises said electron donor moiety.

15. A method according to claim 12, wherein both of said electron donor and acceptor moieties are attached via polydentate nucleosides.

16. A method according to claim 12, further comprising a transition metal chelated to said polydentate nucleoside.

17. A method according to claim 12, wherein the other of said electron donor and acceptor moieties is an electrode.

18. A method according to claim 12 wherein at least one of said electron donor moiety and said electron acceptor moiety is an organic electron donor or acceptor.

19. A method for making a nucleic acid with an electron transfer moiety attached via a polydentate ligand, said method comprising forming a nucleic acid from phosphoramidite nucleosides, at least one of which comprises a polydentate ligand attached to the ribose of said nucleoside.

20. A method according to claim 19 wherein said polydentate ligand further comprises a bound transition metal.