Nucleic acid inhibitors of glutamate receptors
AssigneesCornell University, The Research Foundation of State University of New York
InventorsLi Niu, Zhen Huang, Hua Shi, John Lis
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Abstract
The present invention relates to novel nucleic acid ligands or aptamers that bind to and inhibit the activation of the α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) subtype of ionotropic glutamate receptors. Also disclosed is a novel combination of technologies, i.e., SELEX and laser pulse photolysis for the selection and screening of aptamers that inhibit receptor function and are useful therefore, in the treatment of diseases associated with excessive activation of ionotropic glutamate receptors.
Claims30
An isolated nucleic acid that binds to a glutamate receptor.
A method for identifying nucleic acids that modulate glutamate receptor function, the method comprising:
- a) screening a nucleic acid library for a nucleic acid that binds to a glutamate receptor;
- b) providing a cell that has been transfected to overexpress said glutamate receptor;
- c) measuring glutamate-induced whole-cell current in the cell in the presence and absence of the nucleic acid identified in step a);
- d) comparing the measurement of whole cell current in the presence and absence of nucleic acid, wherein a decrease in the whole-cell current in the cell in the presence of nucleic acid indicates that the nucleic acid inhibits glutamate receptor function.
An isolated DNA that codes for an RNA wherein the RNA comprises a nucleotide sequence selected from the group of SEQ ID NOS.:
- 1 to 11.
In a method for identifying nucleic acid ligands for a receptor of interest, which includes the steps of contacting a mixture of randomized nucleic acids with the desired target, partitioning unbound nucleic acids from those nucleic acids which have bound to the target molecule, dissociating the nucleic acid-target complexes, amplifying the nucleic acids dissociated from the nucleic acid-target complexes to yield a ligand-enriched mixture of nucleic acids, then reiterating the steps of binding, partitioning, dissociating and amplifying through as many cycles as desired to yield high affinity nucleic acid ligands to the target molecule, the improvement consisting of removing unbound ligand by filtering the mixture of nucleic acid-target complexes through a nitrocellulose filter.
Dependencies are inferred from the claim text.