Metallic solid supports modified with nucleic acids

• US 6,291,188 B1
• Filed: 08/15/2000
• Issued: 09/18/2001
• Est. Priority Date: 06/07/1995
• Status: Expired due to Term

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Meade, Thomas J., Kayyem, Jon F.
• Primary Examiner(s): Houtteman, Scott W.
• Application Number:
  US09/639,311
• Time in Patent Office: 399 Days
• Field of Search: 435/6, 435/7.1, 536/24.3, 427/58
• 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. An array of nucleic acids comprising a solid support having a plurality of regions, each region comprising an electrode comprising a self-assembled monolayer comprising:

a) blocking moieties, having a first end attached to said electrode, wherein said blocking moieties shield nucleic acids from said electrode; and

b) at least one modified nucleic acid, comprising a nucleic acid and a linker moiety having a first and second end, wherein said first end of said linker is attached to said electrode and said second end is attached to said nucleic acid;

wherein at least two different regions comprise different nucleic acids.

2. An array according to claim 1 wherein said first end of said blocking moieties is attached to said electrode via a sulfur linkage.

3. An array according to claim 1 wherein said first end of said linker is attached to said electrode via a sulfur linkage.

4. An array according to claim 1, 2 or 3 wherein each electrode comprises gold.

5. An array according to claim 1 wherein said blocking moieties have the formula:

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said electrode;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups;

n is an integer from 3 to 50; and

X is a terminal group.

6. An array according to claim 5 wherein R¹ and R₂ are hydrogen.

7. An array according to claim 1 wherein said blocking moieties comprise alkyl.

8. An array according to claim 5, 6 or 7 wherein n is ≧

6.

9. An array according to claim 1 wherein said blocking moiety is a branched molecule.

10. An array according to claim 1 wherein said linker moiety is a straight chain alkyl group.

11. A straight chain alkyl group according to claim 10 wherein said chain ranges from 1 to 20 carbon atoms.

12. An array according to claim 1 wherein at least two of said blocking moieties are different.

13. An array according to claim 1 wherein at least one of said blocking moieties is a branched molecule.

14. An array according to claim 12 or 13 wherein at least one of said blocking moieties is an alkyl group.

15. An array according to claim 1 wherein said linker moiety has the formula:

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said electrode;

R₁ and R₂ are independently selected from the group consisting of hydrogen and substituent groups;

n is an integer from 3 to 50; and

Y is the point of attachment for a nucleic acid.

16. An array according to claim 15 wherein said linker moiety is a straight chain alkyl group.

17. A straight chain alkyl group according to claim 16 wherein said chain ranges from 1 to 20 carbon atoms.

18. An array according to claim 1 wherein said modified nucleic acids have the formula:

wherein

SCM is a sulfur-containing moiety, wherein said sulfur containing moiety is attached to said electrode;

R₁ and R² are independently selected from the group consisting of hydrogen and substituent groups; and

n is an integer from 3 to 50.

19. An array according to claim 18 wherein R₁ and R₂ are hydrogen.

20. An array according to claim 15, 18 or 19 wherein n is ≧

6.

21. An array according to claim 1 wherein said blocking moiety comprises a phosphorus-containing moiety.

22. An array according to claim 1 wherein said nucleic acid is attached to said linker at a 2′

position of a ribose.

23. An array according to claim 1 wherein said nucleic acid is attached to said linker at a 3′

position of a ribose.

24. An array according to claim 1 wherein said nucleic acid is attached to said linker at a base of said nucleic acid.

25. An array according to claim 1 wherein said nucleic acid is attached to said linker at a phosphate linkage of said nucleic acid.

26. An array according to claim 1 wherein said solid support is glass.

27. An array according to claim 1 wherein said solid support is plastic.

28. A method of hybridizing a probe nucleic acid to a target nucleic acid, said method comprising adding a target nucleic acid to an array of probe nucleic acids comprising a solid support having at plurality of regions, each region comprising an electrode comprising a self-assembled mixed monolayer comprising:

a) blocking moieties, having a first end attached to said electrode, wherein said blocking moieties shield nucleic acids from said electrode; and

b) at least one modified nucleic acid, comprising a probe nucleic acid and a linker moiety having a first and second end, wherein said first end of said linker is attached to said solid support and said second end is attached to said nucleic acid;

wherein at least two different regions comprise different probe nucleic acids;

under conditions wherein at least one of said probe nucleic acids and said target nucleic acid will hybridize to form a hybridization complex.

29. A method according to claim 28 wherein said blocking moiety is a branched molecule.

30. A method according to claim 28 wherein at least two of said blocking moieties are different.

31. A method according to claim 28 wherein said blocking moiety is a straight chain alkyl group.

32. A method according to claim 28 wherein said linker moiety is a straight chain alkyl group.

33. An alkyl chain according to claim 30 or 31 wherein said chain ranges form 1 to 20 carbon atoms.

34. A method according to claim 28 wherein said target nucleic acid is labeled.

35. A method according to claim 34 wherein said target nucleic acid is labeled with a flourescent dye.