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US 2025/0137032 A1Published application
Two-component systems for use in cell-free gene expression
OwnerNeha Prashant Kamat, Justin Alexander Peruzziinventors
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Abstract
Provided herein are cell-free systems comprising two-component systems including a histidine kinase and a response regulator. In embodiments, the systems further include synthetic membranes. Also provided are methods for using the systems to detect analytes and synthesize target molecules.
Claims20
A cell-free system comprising a nucleic acid encoding a histidine kinase of a two-component system;
- a nucleic acid encoding a response regulator of the two-component system;
- and at least one nucleic acid encoding a target molecule operably linked to a response regulator-binding promoter.
A method of detecting an analyte in a sample, the method comprising:
- adding to the sample:
- a synthetic membrane and a cell-free system, the cell-free system comprising:
- a nucleic acid encoding a histidine kinase of a two-component system, wherein the histidine kinase binds the analyte;
- a nucleic acid encoding a response regulator for the two-component system;
- and
- at least one nucleic acid encoding a reporter molecule operably linked to a response regulator-binding promoter;
- and
- detecting the reporter molecule.
5 more elements
A method of synthesizing a target molecule in a sample, the method comprising:
- adding to the sample:
- a synthetic membrane, and a cell-free system, the cell-free system comprising:
- a nucleic acid encoding a histidine kinase of a two-component system;
- a nucleic acid encoding a response regulator of the two-component system;
- and
- at least one nucleic acid encoding a target molecule operably linked to a response regulator-binding promoter;
- and
- adding an activator to the sample, wherein the histidine kinase binds the activator.
5 more elements
Dependencies are inferred from the claim text.