Methods of generating nanoarrays and microarrays
The methods described herein provide a means of producing an array of spatially separated proteins. The method relies on covalently attaching each protein of the plurality of proteins to a structured nucleic acid particle (SNAP), and attaching the SNAPs to a solid support.
1. A method for attaching a protein to a solid support, comprising
(a) providing a seed oligonucleotide, wherein the seed oligonucleotide is covalently attached to a protein;
(b) hybridizing the seed oligonucleotide to an immobilized oligonucleotide, wherein the immobilized oligonucleotide is covalently attached to a solid support, wherein the immobilized oligonucleotide has a region of internal complementarity that is self-hybridizing; and
(c) growing the seed oligonucleotide by polymerization, thereby producing a structured nucleic acid particle, wherein the structured nucleic acid particle comprises a self-hybridized region of internal complementarity, wherein the protein is attached to the solid support via the structured nucleic acid particle.
2. The method of claim 1 , wherein step (b) further comprises covalently attaching the seed oligonucleotide to the immobilized oligonucleotide.
3. The method of claim 2 , wherein the seed oligonucleotide is covalently attached to the immobilized oligonucleotide by a conjugation reaction.
4. The method of claim 1 , wherein the solid support is a particle.
5. The method of claim 4 , wherein the immobilized oligonucleotide is covalently attached to the particle.
6. The method of claim 4 , wherein the protein is attached to the particle via the structured nucleic acid particle.
7. The method of claim 1 , wherein the immobilized oligonucleotide is covalently attached to an attachment site in an array of attachment sites, wherein the attachment site is located on the solid support.
8. The method of claim 7 , wherein the protein is attached to the site via the structured nucleic acid particle.
9. The method of claim 8 , wherein the array comprises a plurality of proteins spatially separated by a distance between 50 nm and 1 um.
10. The method of claim 1 , wherein the structured nucleic acid particle comprises nucleic acid origami.
11. The method of claim 1 , wherein the structured nucleic acid particle is a DNA molecule.
12. The method of claim 11 , wherein the DNA molecule comprises a repeating sequence of nucleotides.
13. The method of claim 1 , wherein the structured nucleic acid particle is a single DNA molecule.
14. The method of claim 1 , further comprising growing the structured nucleic acid particle via a controlled polymerization reaction.
15. The method of claim 1 , wherein the protein is a single protein.
16. The method of claim 1 , wherein the protein forms a protein complex with another protein.
17. The method of claim 1 , wherein the seed oligonucleotide covalently attached to the protein in step (a) is formed by reacting a functional group of the seed oligonucleotide with a functional group of the protein.
18. The method of claim 1 , further comprising removing the structured nucleic acid particle from the solid support.
19. The method of claim 18 , wherein the removing step comprises chemical digestion of the structured nucleic acid particle.
20. The method of claim 18 , wherein the removing step comprises enzymatic digestion of the structured nucleic acid particle.
21. The method of claim 1 , wherein the solid support is in a flow cell.
22. The method of claim 1 , further comprising detecting the protein that is attached to the solid support via the structured nucleic acid particle.
23. The method of claim 1 , further comprising detecting the protein, wherein the detecting step comprises a fluorescence detection method.