Patent ID: 11965205
Assignee: VEDABIO, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method for detecting one or more nucleic acid targets and/or non-nucleic acid targets of interest in a sample comprising the steps of:
providing a reaction mix comprising:
aptamer/aptamer-complement double-strand oligonucleotides;
first ribonucleoprotein complexes (RNP1-NONs), wherein each of the RNP1-NONs comprises a first nucleic acid-guided nuclease and a first gRNA (gRNA1-NON); wherein the gRNA1-NONs comprise a sequence complementary to the aptamer-complement, and wherein the first nucleic acid-guided nuclease exhibits trans-cleavage activity;
second ribonucleoprotein complexes (RNP2s), wherein each of the RNP2s comprises a second nucleic acid-guided nuclease and a second gRNA that is not complementary to the aptamer-complement, and wherein the second nucleic acid-guided nuclease exhibits trans-cleavage activity;
a plurality of blocked nucleic acid molecules each comprising a sequence complementary to the second gRNA;
and a plurality of reporter moieties;, contacting the reaction mix with the sample under conditions that allow non-nucleic acid targets of interest in the sample to bind to the aptamer, wherein:
upon binding of the aptamer to the non-nucleic acid targets of interest, the aptamer-complement is no longer hybridized to the aptamer and the aptamer-complement can bind with one of the RNP1-NONs;
upon binding of the aptamer-complement to one of the RNP1-NONs, an aptamer-complement-bound RNP1-NON becomes active and trans-cleaves at least one of the blocked nucleic acid molecules, thereby producing at least one unblocked nucleic acid molecule that can complex with one of the RNP2s;
upon binding of the at least one unblocked nucleic acid molecule to one of the RNP2s, an unblocked nucleic acid molecule-bound RNP2 becomes active and trans-cleaves at least one more of the blocked nucleic acid molecules and at least one of the reporter moieties in a cascade;
allowing the cascade to continue; and
detecting cleaved reporter moieties, thereby detecting the one or more non-nucleic acid targets of interest in the sample.