Patent ID: 11965204
Assignee: THE HONG KONG UNIVERSITY OF SCIENCE AND TECHNOLOGY
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method of detecting an analyte in a sample of regulating DNA circuits, the method comprising:
a) contacting the sample to a composition comprising a restriction enzyme, a DNA template, a sensor molecule, and an invading probe, wherein the analyte binds to the sensor molecule and permits recognition of a restriction site in the DNA template by the restriction enzyme and wherein the invading probe generates a detectable signal in conjunction with a reporter molecule and
b) binding the sensor molecule to the DNA template; wherein the sensor molecule excludes the restriction enzyme from a restriction site specific to said restriction enzyme in the DNA template in the absence of the analyte or displacing the sensor molecule from the DNA template if the analyte is present and binds to the sensor molecule,
wherein the sensor molecule is an allosteric transcription factor or an aptamer,
wherein the analyte is a ligand that binds to the sensor molecule,
wherein the restriction enzyme is a type IIS restriction enzyme that generates an overhang at the 3′ end or 5′ end of the DNA template and the overhang is at least four nucleotides in length,
wherein the DNA template is a double-stranded DNA sequence that contains at least one restriction site upstream or downstream of at least one operator sequence or a single-stranded DNA sequence that contains restriction site in the loop region of a hairpin structure and a domain complementary to an aptamer molecule, and
wherein the invading probe is a single-stranded DNA sequence that is partially or fully complementary to the DNA template.