Nucleotides with a 3′ AOM blocking group

Patent metadata

Field	Value
Publication number	US-12227802-B2
Application number	US-202318488801-A
Country	US
Kind code	B2
Filing date	Oct 17, 2023
Priority date	Dec 26, 2018
Publication date	Feb 18, 2025
Grant date	Feb 18, 2025

Abstract

Embodiments of the present disclosure relate to nucleotide molecules with a 3′ AOM blocking group. Also provided herein are methods to prepare such nucleotide molecules, and the uses of fully functionalized nucleotides containing the 3′-OH blocking group for sequencing applications.

First claim

What is claimed is:

1. A nucleotide comprising a 2′ deoxyribose having a removable 3′-OH blocking group forming a structure

covalently attached to the 3′-carbon atom of the 2′ deoxyribose, wherein:
each of R 1a , R 1b , R 2a and R 2b is independently H, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl, cyano, or halogen; and
R 3 is C 2 -C 6 alkenyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl and combinations thereof.

2. The nucleotide of claim 1 , wherein each of R 1a and R 1b is H.

3. The nucleotide of claim 1 , wherein each of R 2a and R 2b is independently H, halogen or C 1 -C 6 alkyl.

4. The nucleotide of claim 3 , wherein each of R 2a and R 2b is H.

5. The nucleotide of claim 3 , wherein each of R 2a and R 2b is methyl.

6. The nucleotide of claim 3 , wherein R 2a is H, and R 2b is halogen or C 1 -C 6 alkyl.

7. The nucleotide of claim 1 , wherein R 3 is C 2 alkenyl optionally substituted with one or more substituents independently selected from the group consisting of halogen, C 1 -C 6 alkyl, C 1 -C 6 haloalkyl and combinations thereof.

8. The nucleoside or nucleotide of claim 7 , wherein R 3 is

9. The nucleotide of claim 1 , wherein the 3′-OH blocking group is selected from the group consisting of

10. The nucleotide of claim 9 , wherein the 3′-OH blocking group is

11. The nucleotide of claim 1 , wherein the nucleotide is covalently attached to a detectable label, optionally via a cleavable linker.

12. The nucleotide of claim 11 , wherein the detectable label is covalently attached to a nucleobase of the nucleotide via a cleavable linker.

13. The nucleotide of claim 12 , wherein the cleavable linker comprises an azido moiety, a —O-allyl moiety, or an acetal moiety.

14. The nucleotide of claim 12 , wherein the 3′-OH blocking group and the cleavable linker are removable under the same chemical reaction conditions.

15. The nucleotide of claim 10 , wherein the nucleotide is covalently attached to a detectable label via a cleavable linker.

16. The nucleotide of claim 1 , wherein the nucleotide is a nucleotide triphosphate.

17. An oligonucleotide or polynucleotide comprising the nucleotide of claim 16 incorporated thereof.

18. The oligonucleotide or polynucleotide of claim 17 , wherein the 3′-OH blocking group of the incorporated nucleotide is

19. The oligonucleotide or polynucleotide of claim 17 , wherein the oligonucleotide or polynucleotide is immobilized on a solid support, and the solid support comprises an array of immobilized oligonucleotides or polynucleotides.

20. The oligonucleotide or polynucleotide of claim 18 , wherein the oligonucleotide or polynucleotide is immobilized on a solid support, and the solid support comprises an array of immobilized oligonucleotides or polynucleotides.

Assignees

• Illumina Cambridge Ltd

Inventors

• Francais Antoine
• Cressina Elena
• Culley Adam
• Mariani Angelica
• Wu Xiaolin
• Liu Xiaohai

Classifications

• C07H21/00

  Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids · CPC title

• C07H19/16

  Purine radicals · CPC title

• C07H19/06 Primary

  Pyrimidine radicals · CPC title

• C12Q2523/107

  Chemical cleaving agents · CPC title

• C12Q2525/186

  incorporating a non-extendable or blocking moiety · CPC title