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 .- 17 . (canceled)
18 . A method of forming an array, comprising:
a) coupling a nucleic acid origami to an attachment site of a solid support;
b) coupling to the nucleic acid origami a seed oligonucleotide, wherein the seed oligonucleotide is attached to a single biological entity; and
c) after coupling the seed oligonucleotide to the nucleic acid origami, growing the seed oligonucleotide by a polymerization reaction.
19 . The method of claim 18 , wherein the nucleic acid origami comprises a long nucleic acid strand hybridized to more than one short nucleic acid strand.
20 . The method of claim 19 , wherein a short nucleic acid strand of the more than one short nucleic acid strands hybridizes to two segments of the long nucleic acid strand.
21 . The method of claim 19 , wherein a short nucleic acid strand of the one or more short nucleic acid strands is complementary to one or more short nucleic acid strands.
22 . The method of claim 21 , wherein a short nucleic acid strand of the one or more short nucleic acid strands is complementary to the seed oligonucleotide.
23 . The method of claim 22 , further comprising hybridizing the seed oligonucleotide to the short nucleic acid strand of the one or more short nucleic acid strands.
24 . The method of claim 19 , wherein a short nucleic acid strand of the one or more short nucleic acid strands has one or more segments that are not complementary to the long nucleic acid strand.
25 . The method of claim 24 , wherein a segment of the short nucleic acid strand that is not complementary is at least 10 nucleotides long.
26 . The method of claim 19 , wherein the seed oligonucleotide comprises a nucleotide sequence that is complementary to the long nucleic acid strand.
27 . The method of claim 25 , further comprising hybridizing the seed oligonucleotide to the long nucleic acid strand.
28 . The method of claim 18 , wherein the nucleic acid origami comprises at least 100 short nucleic acid strands.
29 . The method of claim 18 , wherein the nucleic acid origami comprises at least 10 folds.
30 . The method of claim 18 , wherein step (b) comprises coupling the seed oligonucleotide to the nucleic acid origami distal to a region of the nucleic acid origami that contacts the attachment site.
31 . The method of claim 18 , wherein step (b) comprises coupling the seed oligonucleotide to the nucleic acid origami to form a linker that attaches to the biological entity.
32 . The method of claim 31 , wherein the linker comprises a rigid outpost of the nucleic acid origami.
33 . The method of claim 32 , wherein the rigid outpost is distal to a region of the nucleic acid origami that contacts the attachment site.
34 . The method of claim 18 , wherein the attachment site comprises a filamentous molecule.
35 . The method of claim 34 , wherein the filamentous molecule comprises an oligonucleotide.
36 . The method of claim 34 , wherein step (a) comprises coupling the nucleic acid origami to the filamentous molecule.
37 . The method of claim 36 , wherein coupling the nucleic acid origami to the filamentous molecule comprises coupling a single-stranded oligonucleotide of the nucleic acid origami to the filamentous molecule.
38 . The method of claim 18 , wherein the polymerization reaction comprises a chain-growth polymerization reaction.
39 . The method of claim 18 , wherein the polymerization reaction comprises a step-growth polymerization reaction.
40 . The method of claim 18 , further comprising (d) detecting the single biological entity that is attached to the solid support via the nucleic acid origami.
41 . The method of claim 18 , wherein the single biological entity comprises a single protein.
42 . The method of claim 18 , wherein the single biological entity is covalently attached to the seed oligonucleotide.
43 . The method of claim 18 , wherein the nucleic acid origami is non-covalently coupled to the attachment site.
44 . The method of claim 18 , wherein the nucleic acid origami is covalently coupled to the attachment site.