Methods and Devices for Detecting Mechanical Forces Using Cas Nuclease and Guide RNA Complexes

A method of identifying the effect of a test compound to inhibit a receptor binding to a ligand comprising;

• contacting 1) a device comprising:
• a) a surface,
• b) a first strand of a nucleobase polymer linked to an area of the surface,
• c) a second strand of a nucleobase polymer configured to hybridize with the first strand and is hybridized with the first strand forming a duplex, and
• d) a ligand linked to the second strand;
• 2) a test compound;
• and
• 3) a cell comprising a receptor to the ligand applying receptor-mediated tension to the device at the receptor opposite the ligand after the contacting step whereby the receptor binds the ligand providing receptor-mediated tension and the receptor-mediated tension exerts forces exceeding a total force needed to denature the duplex, whereby the first strand no longer hybridizes to the second strand, the first strand becomes a single stranded nucleobase polymer;
• contacting the first strand with a Cas nuclease and guide RNA complex and a single stranded reporter nucleic acid,
• wherein the reporter nucleic acid comprises a fluorescent dye and a quencher conjugated to the reporter nucleic acid in sufficient spatial proximity such that the fluorescent dye is quenched by the quencher;
• wherein the guide RNA hybridizes with the first strand providing a Cas guide RNA and first strand complex,
• wherein the Cas guide RNA and first strand complex cleave the reporter nucleic acid such that the fluorescent dye is no longer in close proximity of the quencher providing a fluorescent signal;
• and
• detecting the fluorescent signal as an indicator of the ability of the test compound to inhibit receptor binding to the ligand in the presence of the test compound.