Polynucleotide encapsulation and preservation using self-assembling membranes
Polynucleotides such as DNA are stored inside vesicles formed from self-assembling membranes. The vesicles may be protocells, liposomes, micelles, colloidosomes, proteinosomes, or coacervates. The vesicles may include surface functionalization to improve polynucleotide encapsulation and/or to bind polynucleotides having specific sequences. Encapsulation in vesicles provides protection for the polynucleotides. Additional protection is provided by addition of one or more stabilizers. The stabilizer may be nucleic-acid stabilizers that stabilize the polynucleotides or may be a protective structural layer around the vesicles such as a layer of silica. A process for stably storing polynucleotides in vesicles and a process for recovering stored polynucleotides from vesicles are both disclosed. The polynucleotides may be used for storage of digital information.
1. A method for concentrating and stably storing polynucleotides comprising:
synthesizing the polynucleotides to have sequences according to an encoding scheme that encodes digital information in a sequence of nucleotide bases of the polynucleotides;
mixing self-assembling vesicle precursors with a solution containing the polynucleotides;
adding a stabilizer, wherein the polynucleotides, the self-assembling vesicle precursors, and the stabilizer form a self-assembling vesicle containing the polynucleotides;
drying the self-assembling vesicle until substantially all moisture is removed;
storing the self-assembling vesicle at room temperature; and
forming a protective encapsulating layer surrounding the self-assembling vesicle, wherein the protective encapsulating layer comprises silicon dioxide or titanium dioxide.
2. The method of claim 1 , wherein the self-assembling vesicle precursors are amphipathic lipids, colloid particles, conjugated protein-polymers, oppositely charged macroions, polyesters, or polysaccharides.
3. The method of claim 1 , wherein the stabilizer is a nucleic-acid stabilizer that associates with and stabilizes the polynucleotides, or a membrane stabilizer that associates with and stabilizes an external surface of the vesicle.
4. The method of claim 3 , wherein the stabilizer is the nucleic-acid stabilizer and the nucleic-acid stabilizer comprises at least one of a salt, a chelator, a polysaccharide, or a sugar alcohol.
5. The method of claim 1 , further comprising mechanically agitating a mixture of the self-assembling vesicle precursors and the solution containing polynucleotides.
6. The method of claim 1 , further comprising:
etching the protective encapsulating layer surrounding the self-assembling vesicle to expose the self-assembling vesicle;
resuspending the self-assembling vesicle in solution;
disrupting a membrane of the self-assembling vesicle; and
purifying the polynucleotides.
7. The method of claim 6 , wherein etching the protective encapsulating layer comprises contacting the self-assembling vesicle with a hydrogen fluoride solution.
8. The method of claim 6 , wherein disrupting the membrane comprises ultrasonication, contacting the membrane with a hydrolase, or contacting with membrane with a chemical that degrades one or more membrane components.
9. The method of claim 6 , further comprising sequencing the polynucleotides and decoding a sequence of nucleotide bases obtained from the sequencing to recover digital information stored in the polynucleotides.
10. The method of claim 1 , wherein drying the vesicle containing the polynucleotides until substantially all moisture is removed comprises drying the vesicle to a powder.
11. The method of claim 1 , further comprising adding functional groups, wherein the functional groups functionalize the internal surface of the vesicle and attract the polynucleotides such that a first concentration of the polynucleotides inside the vesicle formed from the self-assembling vesicle precursors is higher than a second concentration of the polynucleotides in the solution.
12. The method of claim 1 , wherein the stabilizer is a protective structural layer that encapsulates the vesicle.