Patent ID: 11905552
Assignee: THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 8:
9. A biosensor for assaying a target nucleic acid, the biosensor comprising:
a substrate comprising a substrate surface and at least two electrodes for passing current through the substrate, wherein the substrate surface comprises graphene;
a pyrenebutanoic acid (PBA) linker molecule comprising a pyrene ring moiety and a carboxylate moiety, wherein the linker molecule is conjugated to the substrate surface via non-covalent interaction between the pyrene ring moiety and the substrate surface; and
an immobilized RNP-gRNA probe that includes
a ribonucleoprotein (RNP) conjugated to the carboxylate moiety of the linker molecule via covalent bonding, wherein the RNP comprises inactive Cas9 protein, wherein the linker molecule is joined to the Recombination UV C (RuvC) domain of the inactive Cas9 protein, and wherein the RNP is immobilized to the substrate surface via the linker molecule, and
a guide ribonucleic acid (gRNA) comprising a first sequence bound to the RNP and a second sequence capable of binding to the target nucleic acid,

wherein conductivity of the substrate surface is modulated upon binding of target nucleic acid to the gRNA to thereby enable sensing of the target nucleic acid while the RNP-gRNA probe remains attached to the substrate surface.