Nucleic acid-based linkers for detecting and measuring interactions
View Patent ↗The invention provides compositions comprising nucleic acid complexes for use in monitoring binding interactions and in measuring association and/or dissociation kinetics with or without force, detecting analytes, screening aptamers, and encoding/encrypting information. In some instances, the nucleic acid complexes are double-stranded nicked nucleic acids comprising a scaffold nucleic acid hybridized to one or more oligonucleotides. In some instances, a first and/or a second oligonucleotide are linked to moieties that are known to interact with each other or which are suspected of interacting with each other.
1. A nucleic acid complex comprising
a single-stranded scaffold nucleic acid hybridized to a plurality of single-stranded oligonucleotides thereby forming a linear partially double-stranded nucleic acid, wherein a first single-stranded oligonucleotide in the plurality is linked to a first binding partner and a second single-stranded oligonucleotide in the plurality is linked to a second binding partner,
wherein the first and second binding partners have
(i) binding affinity for an analyte that is neither the first nor the second binding partner or
(ii) binding affinity for each other, and
a loop is created in the scaffold nucleic acid when the first and second binding partners either (i) bind to the analyte in the presence of the analyte or (ii) bind to each other.
2. The nucleic acid complex of claim 1 , wherein the first and second binding partners have binding affinity for an analyte that is neither the first nor the second binding partner.
3. The nucleic acid complex of claim 1 , wherein the first and second binding partners have binding affinity for each other.
4. The nucleic acid complex of claim 1 , wherein the first binding partner is a receptor and the second binding partner is a ligand for the receptor.
5. The nucleic acid complex of claim 1 , wherein the first binding partner is an antibody or an antigen-binding antibody fragment and the second binding partner is an antigen bound by the antibody or the antigen-binding fragment.
6. The nucleic acid complex of claim 1 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 10 nucleotides.
7. The nucleic acid complex of claim 1 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are located about equidistant from the center of the scaffold nucleic acid.
8. The nucleic acid complex of claim 1 , wherein the first binding partner is an antibody or an antigen-binding antibody fragment.
9. The nucleic acid complex of claim 1 , wherein the first and second binding partners are antibodies or antigen-binding antibody fragments.
10. The nucleic acid complex of claim 9 , wherein the first and second binding partners are identical to each other.
11. The nucleic acid complex of claim 1 , wherein the first and second binding partners have binding affinity for the same analyte, and each is an antibody or an antigen-binding antibody fragment.
12. The nucleic acid complex of claim 1 , wherein the first and second binding partners bind to each other or to the analyte non-covalently.
13. The nucleic acid complex of claim 1 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 200 nucleotides.
14. The nucleic acid complex of claim 1 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 1000 nucleotides.
15. A nucleic acid complex comprising
a single-stranded scaffold nucleic acid and a plurality of single-stranded oligonucleotides, each of which hybridizes throughout its length to a contiguous nucleotide sequence in the scaffold nucleic acid, wherein a first single-stranded oligonucleotide in the plurality is linked to a first binding partner, and a second single-stranded oligonucleotide in the plurality is linked to a second binding partner,
wherein the first and second binding partners have
(i) binding affinity for an analyte that is neither the first nor the second binding partner or
(ii) binding affinity for each other, and
a loop is created in the scaffold nucleic acid when the first and second binding partners either (i) bind to the analyte in the presence of the analyte or (ii) bind to each other.
16. The nucleic acid complex of claim 15 , wherein each oligonucleotide is complementary to a contiguous sequence in the scaffold nucleic acid.
17. The nucleic acid complex of claim 15 , wherein the first and second binding partners have binding affinity for an analyte that is neither the first nor the second binding partner.
18. The nucleic acid complex of claim 15 , wherein the first and second binding partners have binding affinity for each other.
19. The nucleic acid complex of claim 15 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 10 nucleotides.
20. The nucleic acid complex of claim 15 , wherein the first binding partner is an antibody or an antigen-binding antibody fragment.
21. The nucleic acid complex of claim 15 , wherein the first and second binding partners are antibodies or antigen-binding antibody fragments.
22. The nucleic acid complex of claim 21 , wherein the first and second binding partners are identical to each other.
23. The nucleic acid complex of claim 15 , wherein the first and second binding partners have binding affinity for the same analyte, and each is an antibody or an antigen-binding antibody fragment.
24. The nucleic acid complex of claim 15 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 1000 nucleotides.
25. A nucleic acid complex comprising
a single-stranded scaffold nucleic acid hybridized to a plurality of single-stranded oligonucleotides thereby forming a linear partially double-stranded nucleic acid, wherein a first single-stranded oligonucleotide in the plurality is linked to a first binding partner, a second single-stranded oligonucleotide in the plurality is linked to a second binding partner, and
wherein the first and second binding partners have
(i) binding affinity for a common analyte that is not a nucleic acid or
(ii) binding affinity for each other, and
a loop is created in the scaffold nucleic acid when the first and second binding partners either (i) bind to the analyte in the presence of the analyte or (ii) bind to each other.
26. The nucleic acid complex of claim 25 , wherein the first and second binding partners bind to each other or to the analyte non-covalently.
27. The nucleic acid complex of claim 25 , wherein the first and second binding partners have binding affinity for a common analyte.
28. The nucleic acid complex of claim 25 , wherein the first and second binding partners have binding affinity for each other.
29. The nucleic acid complex of claim 25 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 10 nucleotides.
30. The nucleic acid complex of claim 25 , wherein the first binding partner is an antibody or an antigen-binding antibody fragment.
31. The nucleic acid complex of claim 25 , wherein the first and second binding partners are antibodies or antigen-binding antibody fragments.
32. The nucleic acid complex of claim 31 , wherein the first and second binding partners are identical to each other.
33. The nucleic acid complex of claim 25 , wherein the first and second binding partners have binding affinity for a common analyte, and each is an antibody or an antigen-binding antibody fragment.
34. The nucleic acid complex of claim 25 , wherein the first single-stranded oligonucleotide and the second single-stranded oligonucleotide are separated from each other by at least 1000 nucleotides.