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Details for Patent: 6,344,318

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Details for Patent: 6,344,318

Title: Methods of producing nucleic acid ligands
Abstract:The present invention includes methods for the identification and production of improved nucleic acid ligands based on the SELEX process. Also included are nucleic acid ligands to the HIV-RT protein identified according to the methods described therein.
Inventor(s): Gold; Larry (Boulder, CO), Tuerk; Craig (Boulder, CO)
Assignee: Gilead Sciences, Inc. (Foster City, CA)
Filing Date:Oct 02, 1998
Application Number:09/165,616
Claims:1. A method for preparing a modified nucleic acid ligand having a specific affinity for a given target comprising:

a) identifying a nucleic acid ligand to said target, wherein said nucleic acid ligand has specific affinity for said target;

b) determining the structure of said nucleic acid ligand; and

c) modifying said nucleic acid ligand, wherein said modifying comprises replacing one or more nucleotides with a modified monomer to produce said modified nucleic acid ligand, wherein said modified nucleic acid ligand mimics the structure of said nucleic acid ligand and wherein said modified nucleic acid ligand has a specific affinity to said target.

2. A method for preparing a modified nucleic acid ligand, wherein said modified nucleic acid ligand is a ligand to a given target, comprising:

a) contacting a candidate mixture of nucleic acids with said target, wherein nucleic acids having an increased affinity to the target 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 ligand-enriched mixture of nucleic acids;

d) repeating steps a)-c), as necessary, to identify a nucleic acid ligand of a given target;

e) determining the structure of said nucleic acid ligand; and

f) modifying said nucleic acid ligand, wherein said modifying comprises replacing one or more nucleotides with a modified monomer to produce a modified nucleic acid ligand, wherein said modified nucleic acid ligand mimics the structure of said nucleic acid ligand and wherein said modified nucleic acid ligand has a specific affinity to said target.

3. A method for preparing a small molecule ligand having a specific affinity for a given target, comprising:

a) identifying a nucleic acid ligand to a given target, wherein said nucleic acid ligand has a specific affinity to said target;

b) determining the structure of said nucleic acid ligand; and

c) modifying said nucleic acid ligand, wherein said modifying comprises replacing each nucleotide of said nucleic acid ligand with a modified monomer to produce the small molecule ligand, wherein said small molecule ligand mimics the structure of said nucleic acid ligand and wherein said small molecule ligand has a specific affinity for said target.

4. A method for preparing a small molecule ligand having a specific affinity for a given target, comprising:

a) contacting a candidate mixture of nucleic acids with said target, wherein nucleic acids having an increased affinity to the target 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 ligand-enriched mixture of nucleic acids;

d) repeating steps a)-c), as necessary, to identify a nucleic acid ligand of a given target;

e) determining the three-dimensional structure of said nucleic acid ligand; and

f) modifying said nucleic acid ligand, wherein said modifying comprises replacing each nucleotide of said nucleic acid ligand with a modified monomer to produce said small molecule ligand, wherein said small molecule ligand mimics the structure of said nucleic acid ligand and wherein said small molecule ligand has a specific affinity for said target.

5. A modified nucleic acid ligand having specific affinity for a target, wherein said modified nucleic acid ligand is identified by the method of claim 1.

6. A modified nucleic acid ligand having specific affinity for a target, wherein said modified nucleic acid ligand is identified by the method of claim 2.

7. A small molecule ligand having specific affinity for a target, wherein said small molecule ligand is identified by the method of claim 3.

8. A small molecule ligand having specific affinity for a target, wherein said small molecule ligand is identified by the method of claim 4.
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