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|Title:||Systematic evolution of ligands by exponential enrichment: chemi-SELEX|
|Abstract:||This application provides methods for identifying nucleic acid ligands capable of covalently interacting with targets of interest. The nucleic acids can be associated with various functional units. The method also allows for the identification of nucleic acids that have facilitating activities as measured by their ability to facilitate formation of a covalent bond between the nucleic acid, including its associated functional unit, and its target.|
|Inventor(s):||Gold; Larry (Boulder, CO), Eaton; Bruce (Boulder, CO), Smith; Drew (Boulder, CO), Wecker; Matthew (Boulder, CO), Jensen; Kirk (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Mar 08, 1995|
|Claims:||1. A method for identifying nucleic acid ligands that bind covalently with a target molecule from a candidate mixture of nucleic acids wherein each nucleic acid in said candidate mixture comprises a chemically reactive functional unit, said method comprising: |
a) preparing a candidate mixture of said nucleic acids;
b) contacting said candidate mixture with said target molecule, wherein nucleic acids which bind covalently with said target molecule form nucleic acid-target molecule complexes;
c) partitioning the nucleic acid-target molecule complexes from the remainder of the candidate mixture, whereby nucleic acid ligands that bind covalently with the target molecule are identified.
2. The method of claim 1 wherein after step c), the nucleic acid is amplified and steps b) through c) repeated until a mixture of nucleic acids enriched in sequences that bind covalently with the target molecule is obtained.
3. The method of claim 1 wherein said chemically reactive functional unit is selected from the group consisting of photoreactive groups, active site directed compounds and peptides.
4. The method of claim 1 wherein the target is modified to include a chemical moiety that reacts with the chemically reactive functional unit of the nucleic acid.
5. The method of claim 1 wherein said nucleic acid comprises a fixed region and a randomized region.
6. The method of claim 5 wherein said chemically reactive functional unit is attached to an oligonucleotide hybridized to said fixed region.
7. The method of claim 1 wherein said target is selected from the group consisting of bradykinin, elastase, and HIV-1 Rev.