Molecular programming tools
The present disclosure provides, in some aspects, nucleic acid-based molecular tools that enable the recording of molecular structure and soluble signals as well as the programmed assembly of molecular structures.
1. A primer exchange reaction system, comprising
(a) a first catalytic molecule comprising (i) an unpaired 3′ toehold domain, (ii) a paired domain that comprises a stopper, and (iii) a linkage domain,
(b) a second catalytic molecule comprising (i) an unpaired 3′ toehold domain, (ii) a paired domain that comprises a stopper, and (iii) a linkage domain,
wherein the 3′ toehold domain of the second catalytic molecule binds to a sequence in the paired domain of the first catalytic molecule;
(c) a primer that binds to the 3′ toehold domain of the first catalytic molecule; and
(c) a polymerase having strand displacement activity.
2. The system of claim 1 , wherein the linkage domain of (a) is a loop domain and the linkage domain of (b) is a loop domain.
3. The system of claim 1 , further comprising a third catalytic molecule comprising (i) an unpaired 3′ toehold domain, (ii) a paired domain that comprises a stopper, and (iii) a linkage domain, wherein the 3′ toehold domain of the third catalytic molecule binds to a sequence in the paired domain of the second catalytic molecule.
4. The system of claim 1 , wherein the primer is linked to a detectable molecule.
5. A cell comprising the system of claim 1 .
6. The system of claim 1 , wherein the linkage domain of (a) and the linkage domain of (b) comprise covalently crosslinked nucleotides.
7. The system of claim 1 , wherein the linkage domain of (a) is a stable paired domain and the linkage domain of (b) is a stable paired domain.
8. The system of claim 1 , wherein the stopper is selected from a synthetic non-DNA linker, a three-carbon linkage, adenylation, azine, digoxigenin, cholesteryl-TEG, I-LINKER, 3-cyanovinylcarbazole, and a non-natural nucleotide sequence.
9. The system of claim 1 , wherein the domains of (a)(i) and (a)(ii) comprise tandem repeat sequences.
10. The system of claim 1 , wherein the unpaired 3′ toehold domain of the first and second catalytic molecule is 5-40 nucleotides in length.
11. The system of claim 1 , wherein the primer is 10-50 nucleotides in length.
12. The system of claim 1 , wherein the unpaired 3′ toehold domain of the first and second catalytic molecule is 5-40 nucleotides in length and the primer is 10-50 nucleotides in length.
13. The system of claim 1 , wherein the first catalytic molecule is a catalytic hairpin molecule and the second catalytic molecule is a catalytic hairpin molecule.