Nucleic acid hydrogel via rolling circle amplification

• US 8,715,732 B2
• Filed: 01/05/2010
• Issued: 05/06/2014
• Est. Priority Date: 01/05/2009
• Status: Active Grant

• Current Assignee: Cornell University
• Original Assignee: Cornell University
• Inventors: Luo, Dan, Lee, Jong Bum, Funabashi, Hisakage
• Primary Examiner(s): Schultz, Doug
• Application Number:
  US12/652,707
• Publication Number:
  US 20100189794A1
• Time in Patent Office: 1,582 Days
• Field of Search: None
• US Class Current: 424/484
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/06 Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels composition of ointments, creams or gels A61K47/00
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione

Abstract

Methods and compositions are provided for producing nucleic acid-based compositions. Methods include enzyme catalyzed or nucleic acid polymer conjugation. Compositions include nucleic acid-containing hydrogels which can be elongated via rolling circle amplification. The hydrogels can encapsulate bioactive agents for drug delivery.

Claims

1. A composition comprising a nucleic acid hydrogel, the nucleic acid hydrogel comprising entangled single-stranded nucleic acid molecules, wherein at least a portion of the single-stranded entangled nucleic acid molecules are formed via rolling cycle amplification, and further wherein said nucleic acid hydrogel has a matrix for structural support, wherein said matrix is formed by gelation of said single-stranded entangled nucleic acid molecules and water, wherein the gelation occurs from a mixture consisting essentially of at least one single-stranded entangled nucleic acid molecule and water.

16. The composition nucleic acid hydrogel of claim 1, wherein said single-stranded entangled nucleic acid molecules are not crosslinked.

17. The composition nucleic acid hydrogel of claim 1, wherein a portion of said single-stranded entangled nucleic acid molecules are coding nucleic acid molecules.

18. The composition nucleic acid hydrogel of claim 1, wherein a portion of said single-stranded entangled nucleic acid molecules are non-coding nucleic acid molecules.

19. The composition nucleic acid hydrogel of claim 1, wherein said hydrogel further comprises one or more enzymes capable of macromolecules necessary for protein modification.

20. The composition nucleic acid hydrogel of claim 1, wherein said hydrogel is elastic.

21. The composition nucleic acid hydrogel of claim 1, wherein the hydrogel is capable of recovering a substantial amount of its original shape after passing the structural support is preserved after injection of the hydrogel through a needle.

22. The composition nucleic acid hydrogel of claim 1, further comprising a drug.

2. A composition comprising a nucleic acid hydrogel, the nucleic acid hydrogel comprising:

a first single-stranded nucleic acid molecule, the first single-stranded nucleic acid molecule formed from the amplification of a circular nucleic acid template using a first primer; and

a second single-stranded nucleic acid molecule, the second single-stranded nucleic acid molecule formed from the amplification of the first single-stranded nucleic acid molecule using a second primer,

and further wherein said nucleic acid hydrogel has a matrix for structural support, wherein said matrix is formed by gelation of said single-stranded nucleic acid molecules and water, wherein the gelation occurs from a mixture consisting essentially of at least one first single-stranded nucleic acid molecule, at least one second single-stranded nucleic acid molecule, and water.

7. The composition nucleic acid hydrogel of claim 2, wherein the first primer is complementary to a portion of the circular nucleic acid template.

8. The composition nucleic acid hydrogel of claim 2, wherein the first primer is complementary to the second primer.

9. The composition nucleic acid hydrogel of claim 2, wherein the second primer is complementary to a portion of the first single-stranded nucleic acid molecule.

10. The composition nucleic acid hydrogel of claim 2, wherein the second single-stranded nucleic acid molecule is entangled with the first single-stranded nucleic acid molecule.

11. The composition nucleic acid hydrogel of claim 2, wherein one or both of the first and second single-stranded nucleic acid molecules comprise deoxyribonucleic acid (DNA), ribonucleic acid (RNA), peptide nucleic acid (PNA), morpholino and locked nucleic acid (LNA), glycol nucleic acid (GNA) and/or threose nucleic acid (TNA).

3. A composition comprising a nucleic acid hydrogel, the nucleic acid hydrogel comprising:

a first single-stranded nucleic acid molecule, the first single-stranded nucleic acid molecule formed from the amplification of a circular nucleic acid template using a first primer; and

a second single-stranded nucleic acid molecule, the second single-stranded nucleic acid molecule formed from the amplification of the first single-stranded nucleic acid molecule using a second primer, and

further comprising a third single-stranded nucleic acid molecule, the third single-stranded nucleic acid molecule formed from the amplification of the second single-stranded nucleic acid molecule using a third primer,

and further wherein said nucleic acid hydrogel has a matrix for structural support, wherein said matrix is formed by gelation of at least one first single-stranded nucleic acid molecule, at least one second single-stranded nucleic acid molecule, at least one third single-stranded nucleic acid molecule, and water, wherein the gelation occurs from a mixture consisting essentially of at least one first single-stranded nucleic acid molecule, at least one second single-stranded nucleic acid molecule, at least one third single-stranded nucleic acid molecule, and water.

4. The composition nucleic acid hydrogel of claim 3, wherein the third primer is complementary to a portion of the circular nucleic acid template.

5. The composition nucleic acid hydrogel of claim 3, wherein the third primer is complementary to a portion of the second single-stranded nucleic acid molecule.

6. The composition nucleic acid hydrogel of claim 3, wherein the third single-stranded nucleic acid molecule is entangled with one or both of the first single-stranded nucleic acid molecule and the second single-stranded nucleic acid molecule.

12. A composition comprising a nucleic acid hydrogel, the nucleic acid hydrogel comprising:

a first single-stranded nucleic acid molecule, the first single-stranded nucleic acid molecule formed from the amplification of a circular nucleic acid template using a first primer; and

a second nucleic acid molecule,

and further wherein said nucleic acid hydrogel has a matrix for structural support, wherein the matrix is formed by gelation of said single-stranded nucleic acid molecules and water, wherein the gelation occurs from a mixture consisting essentially of at least one first single-stranded nucleic acid molecule, at least one second nucleic acid molecule, and water.

13. The composition nucleic acid hydrogel of claim 12, wherein the second nucleic acid molecule is single-stranded.

14. The composition nucleic acid hydrogel of claim 12, wherein the second nucleic acid molecule is double-stranded.

15. A nucleic acid hydrogel, comprising:

a first single-stranded nucleic acid molecule, the first single-stranded nucleic acid molecule formed from the amplification of a circular nucleic acid template using a first primer; and

a second nucleic acid molecule;

wherein the first nucleic acid molecule comprises DNA and the second nucleic acid molecule comprises RNA.

23. The nucleic acid hydrogel of claim 15, wherein the first single-stranded nucleic acid molecule is not crosslinked to the second nucleic acid molecule.

24. The nucleic acid hydrogel of claim 15, wherein the hydrogel further comprises one or more enzymes capable of macromolecules necessary for protein modification.

25. The nucleic acid hydrogel of claim 15, wherein the hydrogel is elastic.

26. The nucleic acid hydrogel of claim 15, wherein the first single stranded nucleic acid molecule and the second nucleic acid molecule are entangled.

27. The nucleic acid hydrogel of claim 15, wherein the hydrogel is capable of recovering a substantial amount of its original shape after passing the structural support is preserved after injection of the hydrogel through a needle.

28. The nucleic acid hydrogel of claim 15, further comprising a drug.