Granted Patent B2

US 12,331,352 · App. 17/576,184 · Granted Jun 17, 2025

Inventors: Jingyue Ju (Englewood Cliffs, NJ); Dae Hyun Kim (Northbrook, IL); Jia Guo (Chandler, AZ); Qinglin Meng (Foster City, CA); Zengmin Li (Flushing, NY); Huanyan Cao (Changzhou, CN)

Assignee: THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK

C12Q1/6874C07H19/04C12Q1/6869C12Q1/6876A61K31/70C12Q2523/107C12Q2523/319C12Q2535/101C12Q2535/113C12Q2563/107

Abstract

Claims (23)

1. A deoxyribonucleic acid having attached at a 3′ end thereof, by a phosphodiester bond, a compound having the structure:

wherein the α labeled O atom is the 3′ O atom of the deoxyribonucleic acid, the wavy line represents the remainder of the deoxyribonucleic acid that is 5′ relative to the 3′O, and wherein R′ is a cleavable chemical group.

2. The deoxyribonucleic acid of claim 1 , wherein R′ is a nitrobenzyl group, an allyl group or a methylazido group.

3. The deoxyribonucleic acid of claim 2 , wherein the deoxyribonucleic acid is attached to a solid surface.

4. The deoxyribonucleic acid of claim 1 , wherein R′ is

wherein the wavy line represents the point of attachment to the 3′O atom.

5. The deoxyribonucleic acid of claim 3 , wherein the deoxyribonucleic acid is attached to the solid surface via 1,3-dipolar azide-alkyne cycloaddition chemistry.

6. The deoxyribonucleic acid of claim 3 , wherein the deoxyribonucleic acid is attached to the solid surface via a polyethylene glycol molecule.

7. The deoxyribonucleic acid of claim 5 , wherein the solid surface is azide-functionalized.

8. The deoxyribonucleic acid of claim 1 , wherein the deoxyribonucleic acid is attached to the solid surface via an azido linkage, an alkynyl linkage or biotin-streptavidin interaction.

9. The deoxyribonucleic acid of claim 3 , wherein the solid surface is in the form of a chip, a bead, a well, a capillary tube, a slide, a wafer, a filter, a fiber, a porous media, or a column.

10. The deoxyribonucleic acid of claim 9 , wherein the solid surface is gold, quartz, silica, plastic, glass, diamond, silver, mental, or polypropylene.

11. The deoxyribonucleic acid of claim 3 , wherein 1000 or fewer copies of the deoxyribonucleic acid are bound to the solid surface.

12. The deoxyribonucleic acid of claim 3 , wherein 2×10 7 , 1×10 7 , 1×10 6 or 1×10 4 or fewer copies of the deoxyribonucleic acid are bound to the solid surface.

13. The deoxyribonucleic acid of claim 1 , wherein the R′ is a cleavable chemical group; and wherein the cleavable chemical group is photocleavable or chemical cleavable.

14. The deoxyribonucleic acid of claim 1 , wherein the base have a detectable marker cleavably linked to it via a cleavable linker.

15. The deoxyribonucleic acid of claim 14 , wherein the detectable marker is a dye, a fluorophore, a chromophore, a combinatorial fluorescence energy transfer tag, a mass tag, or an electrophore.

16. The deoxyribonucleic acid of claim 15 , wherein the fluorophore is a cyanine dye, a green fluorescent dye, a red fluorescent dye or a rhodamine dye.

17. The deoxyribonucleic acid of claim 14 , wherein the linker is cleaved by contacting the linker with tris(2-carboxyethyl) phosphine.

18. The deoxyribonucleic acid of claim 17 , wherein the linker is photocleavable or chemical cleavable.

19. The deoxyribonucleic acid of claim 18 , wherein the linker is 2-nitrobenzyl.

20. The deoxyribonucleic acid of claim 14 , wherein the linker is:

wherein the left-hand wavy line represents the point of attachment to the base, or further molecule connecting to the base, and the right hand wavy line represents the point of attachment to a fluorophore or other detectable markers, or a further molecule connecting to the fluorophore or other detectable markers.

Assignments (1)

Continuity (6)

References Cited (400)