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US 12,142,104 B2Granted
Methods and compositions for molecular authentication
AssigneePresident and Fellows of Harvard College
InventorsPeng Yin, Jocelyn Yoshiko Kishi, Thomas E. Schaus, Youngeun Kim
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Abstract
Provided herein, in some embodiments, are molecular authentication methods, systems, and compositions.
Claims19
A method of detecting a target nucleic acid, comprising
- contacting the target nucleic acid with an authenticating identifier composition, wherein the target nucleic acid comprises a first strand comprising domain x* and domain a, and the authenticating identifier composition comprises:
- (a) a second strand comprising domain a*, domain x, domain b*, and a first molecule of a quencher-fluorophore pair, wherein domain a* binds to domain a, and domain x binds to domain x*;
- (b) a third strand comprising a second molecule of the quencher-fluorophore pair, domain b, and domain x*, wherein domain b binds to domain b*;
- and
- (c) a fourth strand comprising domain b and domain x*;
- wherein binding of the second strand to the first strand produces a detectable signal, wherein the detectable signal is an optical signal.
3 more elements
A method of detecting a target nucleic acid, comprising
- (a) contacting the target nucleic acid with a first catalytic hairpin strand that comprises domain b, a loop domain, domain b* and domain a* and a second catalytic hairpin strand that comprises domain c, a loop domain, domain c* and domain b*, wherein domain b binds to domain b*, and domain c binds to domain c*;
- and
- (b) removing the first and second catalytic hairpin strands from the target nucleic acid;
- (c) combining the removed catalytic hairpin strands with (i) a primer comprising domain a, wherein domain a binds to domain a*, (ii) a strand displacing polymerase, and (iii) dNTPs, and producing a key strand comprising domain a, domain b, and domain c;
- and
- (d) authenticating the key strand using a molecular lock composition, wherein the molecular lock composition comprises a lock strand comprising domain a*, domain b*, and domain c*, and wherein binding of the lock strand to the key strand produces a detectable signal, wherein the detectable signal is an optical signal.
3 more elements
A method of detecting a target nucleic acid, comprising
- (a) contacting the target nucleic acid with a first key strand that comprises domain a and domain x and a second key strand that comprises domain b and domain y;
- and
- (b) removing the first and second key strands from the target nucleic acid;
- (c) combining the removed key strands with a molecular lock composition, wherein the molecular lock composition comprises (i) a first lock strand comprising, in a 5′ to 3′ direction, a first molecule of a quencher-fluorophore pair, domain y*, domain b*, domain x*, and domain a*, (ii) a second lock strand comprising, in a 5′ to 3′ direction, domain x and domain b, and (iii) a third lock strand comprising, in a 5′ to 3′ direction, domain y and a second molecule of the quencher-fluorophore pair, wherein domain a binds to domain a*, domain x binds to domain x*, domain b binds to domain b*, and domain y binds to domain y*, wherein binding of the first and second key strands to the first lock strand produces a detectable signal, wherein the detectable signal is an optical signal.
1 more element
Dependencies are inferred from the claim text.