Back to results
US 5,705,344 AGranted
High-throughput screening assay for inhibitors of nucleic acid helicases
AssigneeTularik Inc.
InventorsHeidi Giordano, Michael G. Peterson, Mohanram Sivaraja
Life of this patent
Estimating the term…
Abstract
The invention provides screening assays for identifying an inhibitor of a pathogenic helicase activity, e.g. a helicase derived from a pathogenic infectious organism such as a bacterium, protozoan or fungus. Helicase activity is detected by the solid-phase, preferential capture of retained (non-liberated) single-stranded nucleic acid comprising a detectable label.
Claims17
A method of identifying an agent that modulates the activity of a helicase comprising the following steps in consecutive order:
- a) forming a mixture of a labeled first nucleic acid hybridized to a complementary unlabeled second nucleic acid, a helicase, a nucleoside triphosphate, and a candidate agent, wherein said first and second nucleic acids are both DNA or are both RNA;
- b) incubating said mixture in a reservoir under conditions wherein said helicase would unhybridize said first and second nucleic acids in the absence of said agent;
- c) immobilizing said second nucleic acid on a solid substrate by direct noncovalent binding, wherein said second nucleic acid has a greater affinity for said solid substrate than does the first nucleic acid;
- d) separating any unhybridized labeled first nucleic acids to obtain isolated, immobilized second nucleic acids;
- e) measuring the amount of said label retained on the immobilized second nucleic acid, wherein the amount of said label is indicative of the modulation of helicase activity by said agent.
2 more elements
Dependencies are inferred from the claim text.