Nanoparticle-oligonucleotide hybrid structures and methods of use thereof

• US 9,121,065 B2
• Filed: 08/09/2011
• Issued: 09/01/2015
• Est. Priority Date: 08/09/2010
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Park, So Jung, Chen, Xi-Jun
• Primary Examiner(s): Martinell, James
• Application Number:
  US13/814,652
• Publication Number:
  US 20130171646A1
• Time in Patent Office: 1,484 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: C12N 15/10 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 Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2563/155 Particles of a defined size, e.g. nanoparticles
• CPC Class Codes: C12Q 2565/113 based on agglutination/precipitation

Abstract

The invention relates to hybrid structures comprising an amphiphilic nucleic acid-block co-polymer assembly on the exterior and a nanoparticle core, and methods of use thereof.

Claims

1. A nanoparticle-nucleic acid hybrid structure, comprising a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core.

2. The hybrid structure of claim 1, wherein said nucleic acid is DNA, RNA, artificial nucleic acids or a combination thereof.

3. The hybrid structure of claim 1, wherein said oligonucleotide is an oligonucleotide ranging in size from 5-100 base pairs.

4. The hybrid structure of claim 1, wherein said block-copolymers comprise an oligonucleotide block and a polystyrene block.

5. The hybrid structure of claim 1, wherein said structure enables detection of single base mismatches from complementary strands.

6. The hybrid structure of claim 1, wherein said structure enhances its stability in water.

7. The hybrid structure of claim 6, wherein said nanoparticle is an iron oxide magnetic particle.

8. The hybrid structure of claim 1, wherein said nanoparticle is a gold nanoparticle, a magnetic nanoparticle, a semiconductor nanoparticle, an insulator nanoparticle, a metallic nanoparticle, a carbon black particle, a quantum dot or any combination thereof.

9. The hybrid structure of claim 8, wherein said iron oxide magnetic particle is functionalized via ligand exchange with alpha-carboxyl polystyrene.

10. The hybrid structure of claim 1, wherein the nanoparticle is a magnetic nanoparticle selected from the group consisting of a metal nanoparticle, a metal oxide nanoparticle, a metalloid nanoparticle, a metalloid oxide nanoparticle, or a combination thereof.

11. The hybrid structure of claim 1, wherein said nanoparticle ranges in size from 1-1000 nanometers in diameter.

12. The hybrid structure of claim 1, wherein said structure enables selective nucleic acid binding to complementary nucleic acid under low salt concentration.

13. The hybrid structure of claim 1, wherein said structure enhances the binding affinity to a nucleic acid complementary to said nucleic acid.

14. The hybrid structure of claim 1, wherein said nucleic acid on said structure selectively binds to a complementary nucleic acid by detecting mismatches between said nucleic acid on said hybrid structure and a nucleic acid contacted by said hybrid structure.

15. The hybrid structure of claim 14, wherein said fluorescence molecule is a fluorescein, a cyanine dye, or a combination thereof.

16. The hybrid structure of claim 1, wherein said structure further comprises a fluorescence molecule.

17. A method for facilitating self-assembly of a nanoparticle-nucleic acid hybrid structure, comprising a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core, the method comprising the step of mixing a pre-determined amount of nucleic acid amphiphilic block-copolymer with a pre-determined amount of nanoparticles to facilitate the self-assembly of said hybrid structure.

18. A method for transfecting a cell with a nucleic acid, comprising contacting a cell with a nanoparticle-nucleic acid hybrid structure, comprising a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core.

19. The method of claim 18, wherein the hybrid structure further comprises a targeting agent.

20. The method of claim 19, further comprising contacting the cell with the hybrid structure in the presence of an applied magnetic field.

21. A method of separating nucleic acid molecules, said method comprising the step of contacting a nucleic said molecule complementary to said nucleic acid molecule comprised by a nanoparticle-nucleic acid hybrid structure, comprising a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core.

22. The method of claim 21, further comprising the step of contacting the nucleic acid with the hybrid structure in the presence of an applied magnetic field.

23. A method for imaging a cell, comprising:

contacting a cell with a nanoparticle-nucleic acid hybrid structure to provide a labeled cell, wherein said nanoparticle-nucleic acid hybrid structure comprises a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core; and

imaging the labeled cell.

24. The method of claim 23, wherein imaging comprises optical imaging, wherein said optical imaging is fluorescence imaging, scattering imaging, colorimetric imaging, electron microscopy imaging, or magnetic resonance imaging.

25. The method of claim 23, wherein the hybrid structure comprises a quantum dot and the imaging comprises fluorescent imaging.

26. The method of claim 23, wherein the hybrid structure comprises a magnetic nanoparticle and the imaging comprises magnetic resonance imaging.

27. The method of claim 23, wherein the hybrid structure further comprises a targeting agent.

28. A method for detecting a complementary nucleic acid sequence with high selectivity, the method comprising the step of utilizing a nanoparticle-nucleic acid hybrid structure in an assay to selectively detect said complementary nucleic acid, wherein said nanoparticle-nucleic acid hybrid structure comprises a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core.

29. The method of claim 28, wherein said selection process is a nucleic acid binding assay that detects binding of said nucleic acid to a complementary nucleic acid.

30. The method of claim 28, wherein said hybrid structure enables detection of a complementary nucleic acid under low salt concentrations.

31. The method of claim 28, wherein said complementary nucleic acid sequence is a sequence of a gene.

32. A method for delivering a composition to a cell, the method comprising:

contacting said cell with a nanoparticle-nucleic acid hybrid structure that comprises a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core.

33. A nanoparticle-nucleic acid hybrid structure, comprising a high density oligonucleotide-amphiphilic block co-polymer exterior assembly and a hydrophobic nanoparticle core, wherein said hybrid structure comprises more oligonucleotides than a structure having said nanoparticle alone without said oligonucleotide-amphiphilic block co-polymer exterior assembly.