DNA origami devices

Patent metadata

Field	Value
Publication number	US-9765341-B2
Application number	US-201113882753-A
Country	US
Kind code	B2
Filing date	Nov 4, 2011
Priority date	Nov 4, 2010
Publication date	Sep 19, 2017
Grant date	Sep 19, 2017

Abstract

Provided herein are DNA origami devices useful in the targeted delivery of biologically active entities to specific cell populations.

First claim

We claim:

1. A DNA origami device comprising a scaffold strand and a plurality of staple strands,
one of the staple strands comprises an aptamer domain capable of binding to an antigen;
another of the staple strands comprises a latch domain hybridized to the aptamer domain, the latch domain sequence selected such that the aptamer domain is capable of binding to the antigen such that the antigen displaces the latch domain;
the aptamer domain and the latch domain, when hybridized to one another, hold the device in a closed configuration; and
the device transitions to an open configuration when the aptamer domain and the latch domain are not hybridized to one another, the DNA origami device has a shape that (a) allows a particle smaller than the inner cross-section of the DNA origami device to access an inner surface of the DNA origami device, and (b) sterically precludes a particle larger than the inner cross-section of the DNA origami device from accessing an inner surface of the device.

2. A DNA origami device comprising a scaffold strand and a plurality of staple strands, wherein:
one of the staple strands comprises a first aptamer domain capable of binding to a first antigen;
another of the staple strands comprises a second aptamer domain capable of binding to a second antigen;
yet another of the staple strands comprises a first latch domain hybridized to the first aptamer domain, the first latch domain sequence selected such that the first aptamer domain is capable of binding to the first antigen such that the first antigen displaces the first latch domain;
still another of the staple strands comprises a second latch domain hybridized to the second aptamer domain, the second latch domain sequence selected such that the second aptamer domain is capable of binding to the second antigen with a greater affinity than it is capable of binding to the second latch domain;
the first aptamer domain hybridized to the first latch domain, and the second aptamer domain hybridized to the second latch domain, hold the device in a closed configuration; and
the device transitions to an open configuration when the first aptamer domain is not hybridized to the first latch domain and the second aptamer domain is not hybridized to the second latch domain, the DNA origami device has a shape that (a) allows a particle smaller than the inner cross-section of the DNA origami device to access an inner surface of the DNA origami device, and (b) sterically precludes a particle larger than the inner cross-section of the DNA origami device from accessing an inner surface of the device.

3. The DNA origami device of claim 1 , wherein the device is biased to the open configuration.

4. The DNA origami device of claim 1 , wherein one of the staple strands comprises a handle domain capable of binding to a moiety.

5. The DNA origami device of claim 4 , wherein multiple staple strands comprise a handle domain capable of binding a plurality of moieties at a stoichiometrically predetermined ratio.

6. The DNA origami device of claim 4 , wherein the plurality of staple strands are selected such that:
a moiety bound by the handle domain is positioned on an inner surface of the DNA origami device when the device is in the closed configuration; and
the transition to the open configuration causes a moiety bound by the handle domain to be positioned on an outer surface of the DNA origami device.

7. The DNA origami device of claim 6 , wherein the handle domain is bound to a moiety.

8. The DNA origami device of claim 7 , wherein the moiety comprises a linker oligonucleotide having a sequence complementary to the sequence of the handle domain, and wherein the moiety is bound to the handle domain through the hybridization of the linker oligonucleotide to the handle domain.

9. The DNA origami device of claim 7 , wherein the moiety comprises at least one of: an antibody, an antibody fragment, a cell surface receptor ligand, a biologically active fragment of a cell surface receptor ligand, a small molecule, a nucleic acid, a DNAzyme, an aptamer, a lipid, a glycan, a glycoprotein, a glycolipid, a proteoglycan, a nanoparticle, a quantum dot, a fluorophore, and a nanocrystal.

10. The DNA origami device of claim 6 , wherein one of the staple strands comprises a second handle domain capable of binding to a second moiety, wherein staple strands are selected such that:
a moiety bound by the second handle domain is positioned on an inner surface of the DNA origami device when the device is in the closed configuration; and
the transition to the open configuration causes a moiety bound by the second handle domain to be positioned on an outer surface of the DNA origami device.

11. The DNA origami device of claim 6 , wherein at least one antigen is a cancer cell-specific antigen.

12. The DNA origami device of claim 1 , wherein staple strands are selected such that the DNA origami device comprises a first domain and a second domain, wherein a first end of the first domain is attached to a first end of the second domain by at least one single-stranded DNA hinge and the second end of the first domain is attached to the second end of the second domain by the hybridization of the aptamer domain to the latch domain.

13. The DNA origami device of claim 12 , wherein staple strands are selected such that the second end of the first domain becomes unattached to the second end of the second domain if the aptamer domain is contacted by a respective antigen.

14. The DNA origami device of claim 2 , wherein the first antigen and the second antigen are different from one another.

15. A DNA origami device comprising a scaffold strand and a plurality of staple strands, wherein:
one of the staple strands comprises a handle domain bound to a moiety; and
staple strands are selected such that:
the DNA origami device has a shape that (a) allows a particle smaller than the inner cross-section of the DNA origami device to access an inner surface of the DNA origami device, and (b) sterically precludes a particle larger than the inner cross-section of the DNA origami device from accessing the inner surface; and
the moiety bound by the handle domain is positioned on the inner surface.

16. A method of delivering a moiety to a cell expressing an antigen, comprising contacting the cell with the DNA origami device of claim 1 carrying the moiety, thereby allowing the antigen to displace the latch domain from the aptamer domain, causing the device to transition to the open configuration, and allowing the moiety to contact the cell.

17. A method of sequestering a particle smaller than the inner cross-section of the DNA origami device in size, comprising contacting the particle with a DNA origami device of claim 1 carrying a moiety, wherein the moiety is capable of specifically binding to the particle.

18. The method of claim 17 , further comprising delivering the sequestered particle to a cell expressing an antigen, comprising contacting the cell with the DNA origami device in which the particle has been sequestered, thereby allowing the antigen to displace the latch domain from the aptamer domain, causing the device to transition to the open configuration and allowing the particle to contact the cell.

19. A DNA origami device comprising a scaffold strand and a plurality of staple strands,
one of the staple strands comprises an aptamer domain capable of binding to an antigen;
another of the staple strands comprises a latch domain hybridized to the aptamer domain, the latch domain sequence selected such that the aptamer

Assignees

• Bachelet Ido
• Douglas Shawn
• Church George
• Harvard College

Inventors

• Bachelet Ido
• Douglas Shawn
• Church George

Classifications

• C12Q2525/205

  Aptamer · CPC title

• C12N2320/32

  Special delivery means, e.g. tissue-specific · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• C12N15/115 Primary

  Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith {; Nucleic acids binding to non-nucleic acids, e.g. aptamers} · CPC title

• A61K48/0025

  wherein the non-active part clearly interacts with the delivered nucleic acid · CPC title