Lipid mediated nucleic acid synthesis

• US 7,772,390 B1
• Filed: 07/18/2007
• Issued: 08/10/2010
• Est. Priority Date: 07/18/2006
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Deamer, David W.
• Primary Examiner(s): Lewis; Patrick T
• Application Number:
  US11/880,111
• Time in Patent Office: 1,119 Days
• Field of Search: None
• US Class Current: 536/25.3
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

Methods and compositions for synthesizing polymers, such as nucleic acids, are provided. Aspects of the invention include methods in which lipids and monomeric precursors, e.g., mononucleotides, of the desired polymeric products are combined to produce a reaction mixture. The reaction mixture is then subjected to one or more steps of drying and rehydrating to produce a desired polymeric product, e.g., nucleic acid. Also provided are the polymeric products themselves, e.g., nucleic acid products, as well as systems and kits for practicing embodiments of the invention.

Claims

1. A method of synthesizing a nucleic acid, said method comprising:

(a) combining a mononucleotide composition and a lipid composition to produce a fluid reaction mixture;

(b) drying said reaction mixture to produce a dried reaction mixture; and

(c) rehydrating said reaction mixture;

to produce a product comprising said nucleic acid.

2. The method according to claim 1, wherein said method comprises at least two iterations of said drying and rehydrating steps.

3. The method according to claim 1, wherein said nucleic acid is a ribonucleic acid.

4. The method according to claim 3, wherein said mononucleotide composition comprises ribonucleotides.

5. The method according to claim 4, wherein said ribonucleotides are chosen from adenosine 5′

-monophosphate and uridine 5′

-monophosphate.

6. The method according to claim 1, wherein said nucleic acid is a homopolymer.

7. The method according to claim 6, wherein said mononucleotide composition is a homogeneous mononucleotide composition.

8. The method according to claim 1, wherein said nucleic acid is a heteropolymer.

9. The method according to claim 8, wherein said mononucleotide composition comprises two or more different types of mononucleotides.

10. The method according to claim 1, wherein said mononucleotide composition and said lipid composition are combined in manner sufficient to provide for a mononucleotide to lipid molar ratio in said reaction mixture that ranges from 2:

1 to 1;

2.

11. The method according to claim 1, wherein said drying comprises subjecting said reaction mixture to a stream of dry gas.

12. The method according to claim 11, wherein said dry gas is carbon dioxide.

13. The method according to claim 1, wherein said rehydrating comprises contacting said dried reaction mixture with an aqueous fluid.

14. The method according to claim 13, wherein said aqueous fluid is an acidic fluid.

15. The method according to claim 14, wherein said acidic fluid is 1 mM HCl.

16. The method according to claim 1, wherein said method further comprises separating product nucleic acids from lipids.

17. The method according to claim 1, wherein said nucleic acid ranges in length from about 40 nt to about 100 nt or longer.

18. The method according to claim 1, wherein said nucleic acid is linear.

19. The method according to claim 1, wherein said method does not employ activated nucleotides.

20. A nucleic acid composition produced according to the method of claim 1, wherein said composition comprises nucleic acid polymers encapsulated in lipid vesicles and said nucleic acid polymers comprise 2′

-5′

linkages that are not recognized by RNase 1.