Aptamer regulated nucleic acids and uses thereof

Patent metadata

Field	Value
Publication number	US-9309568-B2
Application number	US-201414287824-A
Country	US
Kind code	B2
Filing date	May 27, 2014
Priority date	Oct 5, 2004
Publication date	Apr 12, 2016
Grant date	Apr 12, 2016

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.

First claim

The invention claimed is:

1. 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. The method of claim 1 , wherein said extending step is performed by polymerase chain reaction (PCR), strand displacement amplification (SDA), rolling circle amplification, ligase chain reaction, nucleic acid sequence-based amplification, or transcription-mediated amplification.

3. The method of claim 1 , wherein said extension products are detected by colorimetric detection, fluorescent detection, chemiluminescence, gel electrophoresis, or oligonucleotide micro array.

4. The method of claim 1 , wherein said extension product is labeled with one or more fluorophores and/or quenchers which alter the fluorescence of said sample.

5. 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.

Assignees

• Smolke Christina D
• Bayer Travis S
• California Inst Of Techn

Inventors

• Smolke Christina D
• Bayer Travis S

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P37/04

  Immunostimulants · CPC title

• C12Q1/686 Primary

  Polymerase chain reaction [PCR] · CPC title

• C12N15/635

  Externally inducible repressor mediated regulation of gene expression, e.g. tetR inducible by tetracyline · CPC title