Single-stranded DNA nanostructures

Patent metadata

Field	Value
Publication number	US-10550145-B2
Application number	US-201615556436-A
Country	US
Kind code	B2
Filing date	Mar 4, 2016
Priority date	Mar 7, 2015
Publication date	Feb 4, 2020
Grant date	Feb 4, 2020

Abstract

The present disclosure relates to nanostructures assembled from nucleic acid consisting of a single strand of DNA rationally-designed to self-assemble into a hairpin loop, helical domains, and locking domains.

First claim

What is claimed is:

1. A nanostructure formed from a single strand of DNA, wherein the nanostructure comprises:
a first layer containing helical domains and locking domains, wherein at least two helical domains of the first layer are separated from each other by a locking domain;
a second layer comprising helical domains and locking domains, wherein at least two helical domains of the second layer are separated from each other by a locking domain; and
loop domains that connect one helical domain to another helical domain and are located along the periphery of the nanostructure,
wherein a locking domain of the first layer is hybridized to a locking domain of the second layer.

2. The nanostructure of claim 1 , wherein the single strand of DNA has a length of 500 nucleotides to 10,000 nucleotides.

3. The nanostructure of claim 2 , wherein the single strand of DNA has a length of 2,000 nucleotides to 5,000 nucleotides.

4. The nanostructure of claim 1 , wherein the helical domains have a length of 10 to 50 nucleotides.

5. The nanostructure of claim 4 , wherein the helical domains have a length of 10 to 30 nucleotides.

6. The nanostructure of claim 1 , wherein the locking domains have a length of 4 to 20 nucleotides.

7. The nanostructure of claim 6 , wherein the locking domains have a length of 5 to 10 nucleotides.

8. The nanostructure of claim 1 , wherein the loop domains have a length of 10 to 100 nucleotides.

9. The nanostructure of claim 8 , wherein the loop domains have a length of 10 to 50 nucleotides.

10. The nanostructure of claim 9 , wherein the loop domains have a length of 20 nucleotides.

11. The nanostructure of claim 1 , wherein the crossing number of the nanostructure is zero and the nanostructure is unknotted.

12. The nanostructure of claim 1 , wherein the nanostructure contains only parallel crossovers.

13. The nanostructure of claim 1 , wherein the nanostructure contains continuous π-π stacking along greater than 50% of the helical domains of the nanostructure.

14. A method of producing the nanostructure of claim 1 , the method comprising incubating the single strand of DNA under conditions that result in the formation of the nanostructure.

15. A method of producing the nanostructure of claim 1 , the method comprising:
(a) combining in a single reaction mixture
(i) a first DNA template and a second DNA template, wherein the templates comprise end sequences that overlap with each other,
(ii) a first primer having a phosphorothioate modification, wherein the first primer binds to the end of the first DNA template that is opposite to the overlapping end sequences,
(iii) a second primer having a phosphate modification, wherein the second primer binds to the end of the second DNA template that is opposite to the overlapping end sequences, and
(iv) polymerase;

(b) performing on the single reaction mixture a nucleic acid amplification reaction, thereby producing amplified DNA;
(c) exposing the amplified DNA to exonuclease digestion, thereby producing a single strand of DNA; and
(d) heating the single strand of DNA to a temperature of 85° C. to 95° C., and then progressively cooling the single strand of DNA to a temperature of 20° C. to 37° C., thereby producing the nanostructure.

Assignees

• Harvard College

Inventors

• Han Dongran
• Myhrvold Cameron
• Yin Peng

Classifications

• C07H1/00

  Processes for the preparation of sugar derivatives · CPC title

• C12Y207/00

  Transferases transferring phosphorus-containing groups (2.7) · 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

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• C12Y301/11003

  Exodeoxyribonuclease (lambda-induced) (3.1.11.3) · CPC title