Granted Patent
B2
US 12,378,560 · App. 17/084,460 · Granted Aug 5, 2025
Sequence multiplicity within spherical nucleic acids
Assignee:
NORTHWESTERN UNIVERSITY
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Abstract
The present disclosure is directed to spherical nucleic acids (SNAs) comprising a nanoparticle core and an oligonucleotide shell attached to the external surface of the nanoparticle core, wherein the oligonucleotide shell comprises a mixture of class A CpG oligonucleotides and class B CpG oligonucleotides. The disclosure also provides methods of using the SNAs for, e.g., regulation of an immune response and gene regulation.
Claims (23)
1. A spherical nucleic acid (SNA) comprising a nanoparticle core and an oligonucleotide shell attached to the external surface of the nanoparticle core, wherein the oligonucleotide shell comprises a mixture of class A CpG oligonucleotides and class B CpG oligonucleotides, wherein the ratio of class A CpG oligonucleotides to class B CpG oligonucleotides in the mixture is about 7:3 and wherein the SNA does not comprise a class C CpG oligonucleotide.
2. The SNA of claim 1 , wherein the oligonucleotide shell consists of a mixture of class A CpG oligonucleotides and class B CpG oligonucleotides.
3. The SNA of claim 1 , wherein the class A CpG oligonucleotides each comprise (i) an internal palindrome sequence containing a CpG motif and (ii) an at least partially phosphorothioated poly (G) sequence at its 5′ and/or 3′ ends.
4. The SNA of claim 1 , wherein the class A CpG oligonucleotide is DNA.
5. The SNA of claim 1 , wherein the class B CpG oligonucleotides each comprise a fully phosphorothioated sequence comprising one or more CpG motifs.
6. The SNA of claim 1 , wherein the class B CpG oligonucleotide is DNA.
7. The SNA of claim 1 , wherein the oligonucleotide shell comprises about 10 to about 150 oligonucleotides.
8. The SNA of claim 1 , wherein each class A CpG oligonucleotide is about 10 to about 50 nucleotides in length.
9. The SNA of claim 1 , wherein each class B CpG oligonucleotide is about 10 to about 50 nucleotides in length.
10. The SNA of claim 1 , further comprising an inhibitory oligonucleotide.
11. The SNA of claim 10 , wherein the inhibitory oligonucleotide is an antagonist oligonucleotide, antisense DNA, small interfering RNA (siRNA), an aptamer, a short hairpin RNA (shRNA), a DNAzyme, or an aptazyme.
12. The SNA of claim 1 , wherein the nanoparticle core is a metallic core, a semiconductor core, an insulator core, an upconverting core, a micellar core, a dendrimer core, a liposomal core, a polymer core, a metal-organic framework core, a protein core, or a combination thereof.
13. The SNA of claim 1 , further comprising an additional agent, wherein the additional agent is a protein, a small molecule, a peptide, or a combination thereof.
14. The SNA of claim 1 , wherein the SNA is from about 1 to about 150 nanometers (nm) in diameter.
15. The SNA of claim 1 , wherein the oligonucleotide shell comprises about 10 to about 50 oligonucleotides.
16. The SNA of claim 1 , wherein the oligonucleotide shell comprises about 75 to about 100 oligonucleotides.
17. The SNA of claim 1 , wherein the oligonucleotide shell comprises about 10 to about 1000 oligonucleotides.
18. The SNA of claim 1 , wherein each class A CpG oligonucleotide is about 15 to about 26 nucleotides in length.
19. The SNA of claim 1 , wherein each class B CpG oligonucleotide is about 18 to about 28 nucleotides in length.
20. A method of inhibiting expression of a gene comprising the step of hybridizing a polynucleotide encoding the gene product with the spherical nucleic acid (SNA) of claim 1 , wherein hybridizing between the polynucleotide and one or more oligonucleotides in the oligonucleotide shell occurs over a length of the polynucleotide with a degree of complementarity sufficient to inhibit expression of the gene product.
21. A method for up-regulating activity of a toll-like receptor (TLR), comprising contacting a cell having the toll-like receptor with the spherical nucleic acid (SNA) of claim 1 , wherein the toll-like receptor is toll-like receptor 9 (TLR9).
22. A method of treating a disorder comprising administering an effective amount of the SNA of claim 1 to a subject in need thereof, wherein the administering treats the disorder.
23. The method of claim 22 , wherein the disorder is cancer, an infectious disease, an autoimmune disease, or a combination thereof.
Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Mar 29, 2021
From: MIRKIN, CHAD A; HUANG, ZIYIN
To: NORTHWESTERN UNIVERSITY
Reel/Frame 055747/0493 →
Continuity (2)
Provisional Application
62927456
· Oct 29, 2019
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