On demand, portable, cell-free molecular sensing platform
AssigneeNorthwestern University
InventorsMichael Christopher Jewett, Julius B. Lucks, Adam D. Silverman, Khalid K. Alam
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Abstract
Disclosed are methods, devices, kits, components, and compositions for detecting a target molecule in a test sample using a cell-free protein synthesis (CFPS) reaction. The methods, devices, kits, components, and compositions may be utilized for detecting target molecules which may include small molecules and/or metabolites of small molecules. The components and compositions used in the disclosed methods, devices, and kits may be dried or lyophilized and may be present or immobilized on a paper substrate, for example, as a paper test article.
Claims21
A method of detecting a target molecule in a biological or environmental sample, the method comprising:
- (i) obtaining a biological or environmental sample which may or may not contain the target molecule,
- (ii) adding the sample to a cell-free protein synthesis (CFPS) reaction comprising:
- (a) a cell extract prepared from one or more host strains, wherein the one or more host strains comprise:
- transcription-translation reagents, energy, cofactor regeneration;
- (b) one or more target molecule metabolizing enzymes;
- and
- (c) a nucleic acid reporter construct encoding a reporter molecule operably linked to a regulatory element that is regulated by a metabolite of the target molecule;
- wherein, in the presence of the target molecule, the one or more target molecule metabolizing enzymes generates the metabolite of the target molecule;
- wherein the metabolite of the target molecule triggers generation of a detectable output via expression of the reporter molecule from the reporter construct, the detectable output comprising a visual, electronic, or optical output.
A method of detecting a target molecule in a biological or environmental sample, the method comprising:
- (i) obtaining a biological or environmental sample which may or may not contain the target molecule;
- (ii) adding the sample to a cell-free protein synthesis (CFPS) reaction comprising:
- (a) a cell extract prepared from one or more host strains, wherein the one or more host strains comprise:
- transcription-translation reagents, energy, cofactor regeneration;
- (b) a nucleic acid reporter construct encoding a reporter molecule operably linked to a regulatory element that is regulated by the target molecule;
- wherein the target molecule triggers generation of a detectable output via expression of the reporter molecule from the reporter construct, the detectable output comprising a visual, electronic, or optical output;
- wherein the transcription-translation reagents comprise a membrane-bound kinase and a soluble DNA-binding response regulator and the membrane-bound kinase regulates the activity of the soluble DNA-binding response regulator.
Dependencies are inferred from the claim text.