Binding-induced DNA nanomachines
View Patent ↗The invention provides a binding-induced DNA nanomachine that can be activated by proteins and nucleic acids. This new type of nanomachine hamesses specific target binding to trigger assembly of separate DNA components that are otherwise unable to spontaneously assemble. Three-dimensional DNA tracks of high density are constructed on gold nanoparticles functionalized with hundreds of single-stranded oligonucleotides and tens of an affinity ligand. A DNA swing arm, free in solution, can be linked to a second affinity ligand. Binding of a target molecule to the two ligands brings the swing arm to AuNP and initiates autonomous, stepwise movement of the swing arm around the AuNP surface. The movement of the swing arm generates hundreds of oligonucleotides in response to a single binding event. The new nanomachines have several unique and advantageous features over DNA nanomachines that rely on DNA self-assembly.
1. A nanomachine comprising:
a nanoparticle;
a first polynucleotide, the first polynucleotide having a first short sequence and a first spacer sequence, the first spacer sequence being conjugated to the nanoparticle and the first short sequence being conjugated to a first ligand;
a second polynucleotide, having a first end and a second end, the first end being conjugated to the nanoparticle and the second end being conjugated to a second ligand;
a third polynucleotide having a second short sequence, a second spacer sequence and a third ligand, the third ligand being conjugated to the second spacer sequence, the second short sequence being complementary to at least a portion of the first short sequence of the first polynucleotide;
a fourth polynucleotide that is complimentary to at least a portion of the second short sequence and the second spacer sequence of the third polynucleotide and being bound to the third polynucleotide; and
a target molecule that binds to the second ligand and the third ligand where, upon binding, the third polynucleotide is brought into proximity of the first polynucleotide such that the fourth polynucleotide is displaced and the first short sequence binds to the complimentary second short sequence, producing an enzymatic cleavage site, which is then cleaved by an enzyme and releasing the first ligand.
2. The nanomachine of claim 1 wherein the nanoparticle is a gold nanopartide.
3. The nanomachine of claim 1 wherein the enzymatic cleavage site is a nicking endonuclease site.
4. The nanomachine of claim 1 wherein the target molecule is a protein or nucleic acid.
5. The nanomachine of claim 4 wherein the protein is streptavidin.
6. The nanomachine of claim 4 wherein the protein is platelet derived growth factor.
7. The nanomachine of claim 5 wherein the second and third ligands are biotin.
8. The nanomachine of claim 6 wherein the second and third ligands are aptamers configured to bind to platelet derived growth factor.
9. The nanomachine of claim 1 wherein the second and third ligands, and the target molecule comprise a nucleic acid wherein the second and third ligands are complimentary to at least a portion of the target molecule.
10. The nanomachine of claim 1 wherein the first ligand is an effector molecule.
11. The nanomachine of claim 10 wherein the effector molecule is a labeling molecule, an affinity tag, a chemotherapeutic, a cytotoxic agent, an active peptide, a contrast agent, a radiolabel, DNA, or a small molecule inhibitor.
12. A method for effector molecule delivery comprising:
providing a functionalized nanoparticle having a first polynucleotide, the first polynucleotide having a first short sequence and a first spacer sequence, the first spacer sequence being conjugated to the nanoparticle and the first short sequence being conjugated to a first ligand; a second polynucleotide, having a first end and a second end, the first end being conjugated to the nanoparticle and the second end being conjugated to a second ligand; a third polynucleotide having a second short sequence, a second spacer sequence and a third ligand, the third ligand being conjugated to the second spacer sequence, the second short sequence being complementary to at least a portion of the first short sequence of the first polynucleotide; and a fourth polynucleotide that is complimentary to at least a portion of the second short sequence and the second spacer sequence of the third polynucleotide and being bound to the third polynucleotide;
adding a target molecule to the functionalized nanoparticle such that the target molecule binds to the second and third ligands, thereby bringing the third polynudeotide into proximity of the first polynucleotide such that the fourth polynucleotide is displaced and the first short sequence binds to the complimentary second short sequence, producing an enzymatic cleavage site;
enzymatically cleaving the first and third polynucleotides at the enzymatic cleavage site; and
releasing the first ligand, wherein the first ligand is the effector molecule.
13. The method of claim 12 wherein the nanoparticle is a gold nanoparticle.
14. The method of claim 12 wherein the enzymatic cleavage site is a nicking endonuclease site.
15. The method of claim 12 wherein the target molecule is a protein or nucleic acid.
16. The method of claim 15 wherein the protein is streptavidin and the second and third ligands are biotin.
17. The method of claim 15 wherein the protein is platelet derived growth factor and the second and third ligands are aptamers configured to bind to platelet derived growth factor.
18. The method of claim 12 wherein the second and third ligands, and the target molecule comprise a nucleic acid wherein the second and third ligands are complimentary to at least a portion of the target molecule.
19. The method of claim 12 wherein the effector molecule is a labeling molecule, an affinity tag, a chemotherapeutic, a cytotoxic agent, an active peptide, a contrast agent, a radiolabel, DNA, or a small molecule inhibitor.