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Details for Patent: 6,596,491

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Details for Patent: 6,596,491

Title: Tenascin-C nucleic acid ligands
Abstract:Methods are described for the identification and preparation of nucleic acid ligands to tenascin-C. Included in the invention are specific RNA ligands to tenascin-C identified by the SELEX method. Further included in the invention are methods for detecting the presence of a disease condition in a biological tissue in which tenascin-C is expressed.
Inventor(s): Hicke; Brian (Boulder, CO), Warren; Stephen (Boulder, CO), Parma; David (Boulder, CO), Gold; Larry (Boulder, CO)
Assignee: Gilead Sciences, Inc. (Foster City, CA)
Filing Date:May 14, 2001
Application Number:09/854,662
Claims:1. A complex for use in in vivo diagnostics comprising a tenascin-C nucleic acid ligand and a marker.

2. The complex of claim 1 wherein said tenascin-C nucleic acid ligand is identified by the method comprising: a) contacting a candidate mixture of nucleic acids with tenascin-C, wherein nucleic acids having an increased affinity to tenascin-C relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; b) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; c) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to tenascin-C, whereby a nucleic acid ligand of tenascin-C may be identified.

3. The complex of claim 2 wherein said candidate mixture of nucleic acids is comprised of single stranded nucleic acids.

4. The complex of claim 3 wherein said single stranded nucleic acids are ribonucleic acids.

5. The complex of claim 4 wherein said candidate mixture of nucleic acids comprises 2'-F (2'-fluoro) modified ribonucleic acids.

6. The complex of claim 1 wherein said marker is selected from the group consisting of radionuclides, fluorophores, magnetic compounds, and biotin.

7. The complex of claim 6 wherein said radionuclide is selected from the group consisting of technetium-99m (Tc-99m), Re-188, Cu-64, Cu-67, F-18, .sup.125 I, .sup.131 I, .sup.32 P, .sup.186 Re.

8. The complex of claim 7 wherein said marker is technetium-99m.

9. The complex of claim 8 further comprising a linker.

10. The complex of claim 9, wherein said linker has the structure ##STR1##

11. The complex of claim 10, wherein said complex is ##STR2##

12. A method for the preparation of a complex comprised of a tenascin-C nucleic acid ligand and a marker, said method comprising: a) contacting a candidate mixture of nucleic acids with tenascin-C, wherein nucleic acids having an increased affinity to tenascin-C relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; b) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; c) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to tenascin-C to yield a tenascin-C nucleic acid ligand; and d) covalently linking said tenascin-C nucleic acid ligand with a marker.

13. The method of claim 12 wherein said marker is selected from the group consisting of radionuclides, fluorophores, magnetic compounds, and biotin.

14. The method of claim 13 wherein said radionuclide is selected from the group consisting of Tc-99m, Re-188, Cu-64, Cu-67, F-18, .sup.125 I, .sup.131 I, .sup.32 P, .sup.186 Re.

15. The method of claim 14 wherein said marker is Tc-99m.

16. The method of claim 15 further comprising a linker.

17. The complex of claim 16, wherein said linker has the structure ##STR3##

18. The complex of claim 17, wherein said complex is ##STR4##
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