Molecular sensing platform and methods of use

Abstract

Provided are signal-inducing CRISPR-sensitive nucleic acid, optionally DNA, sensors, for example comprising: a) a non-functional CRISPR-sensitive DNA reporter construct comprising a non-functional expression cassette with at least one CRISPR target site inserted, generated by removal or addition of nucleic acids or naturally present in the expression cassette, the non-functional expression cassette having a reporter construct upstream end upstream of the CRISPR target site and a reporter construct downstream end downstream of the CRISPR target site, and b) a function-restoring nucleic acid, the function-restoring nucleic acid comprising an upstream flanking end, a function restoring repair insert and a downstream flanking end, wherein the upstream flanking end interfaces with reporter construct upstream end and/or the downstream flanking end interfaces with the reporter construct downstream end and one or both of the flanking ends are capable of permitting insertion or ligation of the function restoring repair insert into/to the reporter construct when the CRISPR target site is actuated under sensing condition, thereby producing a functional DNA reporter construct and sensor signal. Also provided are cell free and cell based systems, kits, primer pairs and molecular barcodes and methods of use thereof.

Dependencies are inferred from the claim text.