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US 9,309,568 B2Granted
Aptamer regulated nucleic acids and uses thereof
AssigneeCalifornia Institute of Technology
InventorsChristina D. Smolke, Travis S. Bayer
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Abstract
The invention relates to aptamer-regulated, ligand-responsive nucleic acids, or “ampliSwitches,” and uses thereof. Particular embodiments include a ligand-responsive nucleic acid that comprises a primer sequence domain and an aptamer domain that is responsive to a ligand.
Claims5
A method for detecting a ligand or analyte in a sample, comprising:
- forming a mixture comprising said sample, a target template, and a nucleic acid comprising (i) a priming sequence that hybridizes to said target template to form a primer:template pair, and (ii) an aptamer that binds to said ligand, if said ligand is present, wherein binding of said ligand to said aptamer causes a conformational change in the nucleic acid that allows said priming sequence to hybridize to said target template;
- incubating said mixture under conditions which allow said priming sequence to hybridize to said target template;
- extending said primer sequence to synthesize an extension product that is complementary to said target template;
- and
- detecting said extension products, wherein the presence of said extension products is indicative of detection of said ligand.
2 more elements
A method for detecting two or more different ligands in a sample, comprising:
- forming a mixture comprising said sample, two or more unique target templates, and a library of two or more different nucleic acids each comprising (i) a unique priming sequence that hybridizes to one of said unique target templates to form a primer:template pair, and (ii) a unique aptamer that binds to one of said different ligands, if present, wherein binding of one of said different ligands to said unique aptamer causes a conformational change in the nucleic acid and allows said unique priming sequence to hybridize to one of said unique target templates;
- incubating said mixture under conditions which allow each said unique priming sequence to hybridize to one of said unique target templates;
- extending each said priming sequence to synthesize an extension product that is complementary to one of said unique target templates;
- detecting said extension product;
- and
- associating the presence of said extension product with one of said different ligands, thereby detecting said two or more different ligands in the sample.
3 more elements
Dependencies are inferred from the claim text.