Compositions and methods relating to complex nucleic acid nanostructures

Patent metadata

Field	Value
Publication number	US-9975916-B2
Application number	US-201314440907-A
Country	US
Kind code	B2
Filing date	Nov 6, 2013
Priority date	Nov 6, 2012
Publication date	May 22, 2018
Grant date	May 22, 2018

Abstract

The invention provides SST motifs of controlled size and shape, comprised of a plurality of oligonucleotides, and methods for their synthesis. The motifs are formed, at least in part, by the self-assembly of single stranded oligonucleotides into curved, corrugated or twisted structures. The location of each oligonucleotide in the resultant motif is known. Accordingly, the motifs may be modified with specificity.

First claim

What is claimed is:

1. A nucleic acid structure comprising
a plurality of annealed motifs, wherein each of the motifs comprises four oligonucleotides, each of the oligonucleotides comprises at least four domains arranged into at least two parallel double helices, every domain of an oligonucleotide of the plurality is bound to a domain of another oligonucleotide of the plurality to form a plurality of S-shaped oligonucleotides, and the nucleic acid structure is curved, corrugated or twisted.

2. The nucleic acid structure of claim 1 , wherein each oligonucleotide comprises at least six domains.

3. The nucleic acid structure of claim 1 , wherein the length of each domain of a single oligonucleotide is the same.

4. The nucleic acid structure of claim 1 , wherein the length of at least two domains, at least three domains or at least four domains of a single oligonucleotide is the same.

5. The nucleic acid structure of claim 1 , wherein the length of each domain of a single oligonucleotide is different.

6. The nucleic acid structure of claim 1 , wherein the oligonucleotides are 18-104 nucleotides in length.

7. The nucleic acid structure of claim 1 , wherein the length of the domains of the oligonucleotides range from 4 nucleotides to 13 nucleotides.

8. The nucleic acid structure of claim 1 , wherein the nucleic acid structure is comprised of different types of oligonucleotides.

9. The nucleic acid structure of claim 1 , further comprising a linker between at least two domains.

10. The nucleic acid structure of claim 1 , wherein the single stranded oligonucleotides are DNA oligonucleotides.

11. The nucleic acid structure of claim 1 , wherein the single stranded oligonucleotides are L-DNA oligonucleotides.

12. The nucleic acid structure of claim 1 , wherein the nucleic acid structure further comprises double crossovers.

13. A composition comprising a plurality of nucleic acid structures of claim 1 , wherein the plurality is at least 50% homogeneous.

14. A method of producing the nucleic acid structure of claim 1 , the method comprising
annealing a plurality of single stranded oligonucleotides in a single vessel to form the nucleic acid structure, wherein at least one single stranded oligonucleotide is present at a molar concentration that is at least 10-fold lower than the molar concentration of other oligonucleotides in the plurality.

15. The method of claim 14 , wherein a first subset of oligonucleotides comprises 2 domains and a second subset of oligonucleotides comprises 4 domains or 6 domains.

16. A composite nucleic acid structure comprising at least two nucleic acid structures of claim 1 , conjugated to each other through a linker.

17. The nucleic acid structure of claim 1 , wherein one or more of the oligonucleotide(s) is conjugated to a moiety selected from the group consisting of a metallic, organic, or inorganic moiety.

18. The nucleic acid structure of claim 17 , wherein the moiety is a metallic moiety selected from gold nanoparticles, quantum dots, and carbon nanotubes.

19. The nucleic acid structure of claim 1 , wherein one or more of the oligonucleotide(s) is conjugated to a nucleic acid moiety.

20. The nucleic acid structure of claim 19 , wherein the nucleic acid moiety binds to one or more oligonucleotide(s) in the nucleic acid structure.

21. The nucleic acid structure of claim 19 , wherein the nucleic acid moiety comprises a triplex forming oligonucleotide.

22. The nucleic acid structure of claim 19 , wherein the nucleic acid moiety comprises a region of complementarity to the one or more oligonucleotide(s) in the nucleic acid structure.

23. The nucleic acid structure of claim 1 , wherein one or more of the oligonucleotide(s) is conjugated to a non-nucleic acid moiety.

24. The nucleic acid structure of claim 23 , wherein the non-nucleic acid moiety is attached covalently or non-covalently to the oligonucleotides.

25. The nucleic acid structure of claim 23 , wherein the non-nucleic acid moiety is a protein, peptide, or polysaccharide.

26. The nucleic acid structure of claim 23 , wherein the non-nucleic acid moiety is a growth factor or an extracellular matrix component.

Assignees

• Harvard College

Inventors

• Yin Peng
• Wei Diming
• Dai Mingjie
• Myhrvold Cameron
• Ke Yonggang
• Jungmann Ralf

Classifications

• C12N15/1031

  mutagenesis by gene assembly, e.g. assembly by oligonucleotide extension PCR · CPC title

• C12N15/10

  Processes for the isolation, preparation or purification of DNA or RNA (chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34) · CPC title

• C07H1/00

  Processes for the preparation of sugar derivatives · CPC title

• C07H21/04 Primary

  with deoxyribosyl as saccharide radical · CPC title

• C12Q1/68

  involving nucleic acids · CPC title