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|Title:||Systematic evolution of ligands by exponential enrichment: tissue selex|
|Abstract:||This invention discloses high-affinity oligonucleotide ligands to complex tissue targets, specifically nucleic acid ligands having the ability to bind to complex tissue targets, and the methods for obtaining such ligands. Tissue targets comprise cells, subcellular components, aggregates or cells, collections of cells, and higher ordered structures. Specifically, nucleic acid ligands to red blood cells ghosts, endothelia of the blood brain and CSF-blood barriers, glioblastomas, and lymphomas are described.|
|Inventor(s):||Heilig; Joseph S. (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||Gilead Sciences, Inc. (Foster City, CA)|
|Filing Date:||Feb 14, 2002|
|Claims:||1. A method for identifying nucleic ligands to target molecules within a blood brain barrier or CSF-blood barrier tissue comprising: a) preparing a candidate mixture of nucleic acid sequences; b) contacting said candidate mixture of nucleic acids with said blood brain barrier or CSF-blood barrier tissue, wherein nucleic acids having an increased affinity to the target molecules relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; c) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and d) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acid sequences with relatively higher affinity and specificity for binding to said target molecules, whereby nucleic acid ligands of said target molecules may be identified. |
2. The method of claim 1 further comprising: e) repeating steps b), c) and d).
3. The method of claim 1 wherein the tissue is selected from the group consisting of cerebral endothelial cells and choroid plexus epithelial cells.
4. The method of claim 1 wherein said candidate mixture is comprised of single-stranded nucleic acids.
5. The method of claim 4 wherein said single-stranded nucleic acids are ribonucleic acids.
6. The method of claim 4 wherein said single-stranded nucleic acids are deoxyribonucleic acids.
7. A nucleic acid ligand to a blood brain barrier or CSF-blood barrier molecule identified according to the method of claim 1.
8. A purified and isolated non-naturally occurring nucleic acid ligand to a blood brain barrier or CSF-blood barrier tissue.
9. The purified nucleic acid ligand of claim 8 which is a non-naturally occurring nucleic acid ligand having a specific binding affinity for a blood brain barrier or CSF-blood barrier molecule, such molecule being a three dimensional chemical structure other than a polynucleotide that binds to said nucleic acid ligand through a mechanism which predominantly depends on Watson/Crick base pairing or triple helix binding, wherein said nucleic acid ligand is not a nucleic acid having the know physiological function of being bound by the molecule.
10. The nucleic acid ligand of claim 8 which is a deoxyribonucleic acid ligand.
11. The nucleic acid ligand of claim 8 which is a ribonucleic acid ligand.
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