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|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 (Fort Collins, CO), Parma; David (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||Gilead Sciences, Inc. (Foster City, CA)|
|Filing Date:||Jan 28, 2000|
|Claims:||1. A method for detecting the presence of a disease in a biological tissue which may contain said disease, wherein said disease is characterized by the expression of tenascin-C in said tissue and wherein said disease is selected from the group consisting of cancer, psoriasis, and atherosclerosis, the method comprising: a) attaching a marker that can be used in in vivo diagnostics to a tenascin-C nucleic acid ligand to form a marker-nucleic acid ligand complex wherein said tenascin-C nucleic acid ligand is selected from the group consisting of SEQ ID NO: 4 65; b) exposing said biological tissue which may contain said disease to said marker-nucleic acid ligand complex; and c) detecting the presence of said disease in said tissue by detecting the presence of said marker-nucleic acid ligand in said tissue. |
2. The method of 1 wherein said marker is selected from from the group consisting of radionuclides, fluorophores, magnetic compounds, and biotin.
3. The method of 2 wherein said radionuclide is selected from the group consisting of technetium-99m (Tc-99m), Re-188, Cu-64, Cu-67, F-18, .sup.125I, .sup.131I, .sup.111In, .sup.32P, and .sup.186Re.
4. The method of 3 wherein said marker is technetium-99m.
5. The method of 4 wherein said tenascin-C nucleic acid ligand comprises a linker.
6. The method of 5 wherein said linker is (CH.sub.2CH.sub.2O).sub.6.
7. The method of 5, wherein said linker has the structure ##STR00001##
8. The method of 1 wherein said tenascin-C nucleic acid ligand is 5'-B-G667667CG-(CH.sub.2CH.sub.2O).sub.6-CGUCGCCGU77U667U6UUUU6CUCCCU65 wherein: all pyrimidines are 2'F; 6=2'OMe G; 7=2'OMe A; 5=3'--3'dT; and B=linker.
9. The method of 8 wherein said technetium-99m is associated with a chelator.
10. The method of 9, wherein said complex is ##STR00002##
11. The method of 10 wherein said complex is ##STR00003##
12. The method of 1 further comprising attaching a therapeutic or diagnostic agent to said complex.
13. The method of 1 wherein said disease is cancer.
14. The method of 1 wherein said tenascin-C nucleic acid ligand is identified by: i) 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; ii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; iii) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids with relatively higher affinity and specificity for binding to tenascin-C, whereby a nucleic acid ligand of tenascin-C is identified.
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