Rapid fluorescence detection of binding to nucleic acid drug targets labelled with highly fluorescent nuclotide base analogs
AssigneeThe United States of America as represented by the Secretary of Commerce
InventorsJohn P. Marino, James T. Stivers, Karen Lacourciere
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Abstract
The present invention is a method of identifying ligands that interact will cellular processes involved in the life-cycle of HIV. In particular, an oligonucleotide, corresponding to specific RNA sequence within an infected cell, is modified by the substitution of 2-aminopurine. As a result, interactions between the oligonucleotide and the ligand can be measured via fluorescence. This technique can be use to find inhibitors of binding between rev and its response element (RRE), dimerization initiation sequences, and topoisomerases and DNA.
Claims35
A method for monitoring and screening for active drugs comprising:
- providing a nucleic acid target modified by the inclusion of 2-aminopurine;
- introducing a ligand;
- and measuring interactions between the nucleic acid target and the ligand.
An oligonucleotide comprising SEQ ID NO:
- 2.
An oligonucelotide comprising SEQ ID NO:
- 3.
An oligonucleotide comprising SEQ ID NO:
- 4.
An oligonucleotide comprising SEQ ID NO:
- 6.
An oligonucleotide comprising SEQ ID NO:
- 7.
An oligonucleotide comprising SEQ ID NO:
- 8.
An assay for identifying a compound that inhibits the binding of HIV rev to HIV RRE, comprising:
- a ligand;
- rev in the presence of modified RRE, having a stem-loop IIB, including 2-aminopurine;
- and means for determining the change in binding of rev to the modified RRE.
An assay for identifying a compound that inhibits the binding of HIV rev to HIV RRE, comprising:
- a first modified RRE, including 2-aminopurine at position 72 of a stem-loop IIB;
- and a second modified RRE, including 2-aminopurine at positions 68 and 72.
Dependencies are inferred from the claim text.