Non-enzymatic, salt-mediated synthesis of polynucleic acids
Provided herein is a method for synthesizing polynucleic acids, comprising the steps of (a) providing an acidic solution substantially free of nucleic acid polymerase and lipids, but containing mononucleotides and a monovalent salt; (b) drying and resolubilizing the mixture of step (a) a plurality of times; and (c) recovering polynucleic acids from a resolubilized mixture of step (b). In certain aspects, the method further uses a low pH, e.g. about 3; it can utilize monophosphates, such as AMP rather than ATP; and it can be used with a polynucleotide template to form a sequence at least partially complementary to said template. Thus, both single-stranded and double-stranded polynucleic acids are provided. Ammonia salts have been used to obtain RNA lengths from 10 to 300 nucleotides after 16 half hour cycles and an effective temperature includes between 80° C. and 100° C.
1. A composition for preparing a polynucleotide from mononucleotides comprising:
an acidic solution or a dried solid prepared from the acidic solution, free of polymerase and lipids, and comprising:
a monovalent salt, wherein the monovalent salt is a metal halide salt or an ammonium halide salt at a concentration of at least 0.01 M; and
mononucleotides selected from the group consisting of adenosine 5′-monophosphate, uridine 5′-monophosphate, guanosine 5′-monophosphate, and cytidine-5′-monophosphate, or 2′-deoxyadenosine 5′-monophosphate, thymidine 5′-monophosphate, 2′-deoxyguanosine 5′-monophosphate, and 2′-deoxycytidine-5′-monophosphate, each at a concentration of from 0.001M to 3M.
2. The composition of claim 1 , wherein the mononucleotides are selected from the group consisting of adenosine 5′-monophosphate, uridine 5′-monophosphate, guanosine 5′-monophosphate, and cytidine-5′-monophosphate, and wherein the polynucleotide is ribonucleic acid (RNA).
3. The composition of claim 1 , wherein the mononucleotides are selected from the group consisting of 2′-deoxyadenosine 5′-monophosphate, thymidine 5′-monophosphate, 2′-deoxyguanosine 5′-monophosphate, and 2′-deoxycytidine-5′-monophosphate, and wherein the polynucleotide is deoxyribonucleic acid (DNA).
4. The composition of claim 1 , further comprising a template polynucleotide complementary to the polynucleotide.
5. The composition of claim 1 , wherein the composition comprises an acidic solution.
6. The composition of claim 5 , wherein the acidic solution is adjusted to a pH between 2 and 4.
7. The composition of claim 5 , wherein the monovalent salt concentration is between 0.05 and 2M in solution.
8. The composition of claim 5 , wherein the monovalent salt is a metal halide salt.
9. The composition of claim 8 , wherein the metal halide salt is selected from the group consisting of NaF, CsCl, NaBr, NaClO 4 , NaCl, LiCl, and KCl.
10. The composition of claim 9 , wherein the metal halide salt is present in the acidic solution at a concentration between 0.05 M and 2M.
11. The composition of claim 5 , wherein the monovalent salt is an ammonium halide salt.
12. The composition of claim 11 , wherein the ammonium halide salt is NH 4 Cl.
13. The composition of claim 12 , wherein the NH 4 Cl is present in the acidic solution at a concentration between 0.05 M and 2M.
14. The composition of claim 5 , wherein the acidic solution is maintained within a temperature range of 60° C. and 90° C.
15. The composition of claim 1 , wherein the composition comprises a dried solid prepared from the acidic solution.
16. The composition of claim 15 , wherein the monovalent salt is a metal halide salt.
17. The composition of claim 15 , wherein the metal halide salt is selected from the group consisting of NaF, CsCl, NaBr, NaClO 4 , NaCl, LiCl, and KCl.
18. The composition of claim 15 , wherein the monovalent salt is an ammonium halide salt.
19. The composition of claim 18 , wherein the ammonium halide salt is NH 4 Cl.
20. The composition of claim 15 , wherein the dried solid is maintained within a temperature range of 60° C. and 90° C.