Patent ID: 11905552
Assignee: KECK GRADUATE INSTITUTE OF APPLIED LIFE SCIENCES
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 9:
10. 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 interaction between the pyrene ring moiety and the substrate surface; and
an RNP-gRNA probe immobilized to the substrate surface via the linker molecule, the RNP-gRNA probe including
a ribonucleoprotein (RNP) comprising Cas9 protein coupled to a second moiety of the linker molecule via a carbodiimide bond, wherein the carbodiimide bond joins the linker molecule at the Recombination UV C (RuvC) domain of the Cas9 protein, 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.