|Title:||High affinity TGF.beta. nucleic acid ligands|
|Abstract:||Methods are described for the identification and preparation of high-affinity nucleic acid ligands to TGF.beta.. Included in the invention are specific RNA and ssDNA ligands to TGF.beta.1 identified by the SELEX method. Also included are RNA ligands that inhibit the interaction of TGF.beta.1 with its receptor.|
|Inventor(s):||Toothman; Penelope J. (Boulder, CO), Ringquist; Steven (Lyons, CO), Gold; Larry (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Jun 02, 1995|
|Claims:||1. A method of identifying nucleic acid ligands to transforming growth factor beta (TGF.beta.) comprising: |
a) preparing a candidate mixture of nucleic acids;
b) contacting the candidate mixture of nucleic acids with TGF.beta., wherein nucleic acids having an increased affinity to TGF.beta. 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 TGF.beta., whereby nucleic acid ligands of TGF.beta. 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 said candidate mixture of nucleic acids is comprised of single stranded nucleic acids.
4. The method of claim 3 wherein said single stranded nucleic acids are ribonucleic acids.
5. The method of claim 4 wherein said nucleic acids comprise 2'-amino (2'-NH.sub.2) modified ribonucleic acids.
6. The method of claim 4 wherein said nucleic acids comprise 2'-F modified ribonucleic acids.
7. The method of claim 4 wherein said nucleic acids comprise 2'-NH.sub.2 -UTP, 2'-F-CTP modified ribonucleic acids.
8. The method of claim 4 wherein said nucleic acids comprise 2'-F-UTP, 2'-NH.sub.2 -CTP modified ribonucleic acids.
9. The method of claim 3 wherein said single stranded nucleic acids are deoxyribonucleic acids.
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