US20190144482A1 – May 16, 2019 – Reversibly Blocked Nucleoside Analogues And Their Use

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Inventors :

Handong Li; Snezana Drmanac; Radoje Drmanac; Xun Xu; Lingling Peng; Scott Gablenz

Owner :

Complete Genomics, Inc. BGI Shenzhen

Application Number :

US16094845

Document Number :

US20190144482A1

Priority Date :

April 21, 2017

Filing Date :

April 21, 2017

Date of Grant/ Publication :

May 16, 2019

Class :

; C07H15 / 04; C07H19 / 10; C07H21 / 04; C07H19 / 20; C12Q1 / 6869

Abstract

Reversibly blocked nucleoside analogues and methods of using such nucleoside analogues for sequencing of nucleic acids are provided.

Claim(s)

1 . A nucleoside analogue of the following formula: wherein R 1 is a reversible blocking group selected from the group consisting of cyanoethenyl, allenyl, cyanoethyl, formaldehyde oximyl, acrylaldehyde oximyl, propionaldehyde oximyl, cyanoethenaldehyde oximyl, a substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted heteroalkenyl, or substituted or unsubstituted heteroalkynyl; R 2 comprises a nucleobase; and R 3 is a cleavable linking moiety comprising at least three phosphates, or analogues thereof.;
4 . (canceled);
5 . (canceled);
6 . (canceled);
7 . (canceled);
8 . (canceled);
9 . (canceled);
10 . A nucleoside analogue of Formula VI: wherein R 1 is a reversible blocking group selected from the group consisting of cyanoethenyl, allenyl, cyanoethyl, formaldehyde oximyl, acrylaldehyde oximyl, propionaldehyde oximyl, and cyanoethenaldehyde oximyl; R 2 is a nucleobase; L is a linker; A 1 comprises an affinity tag; X is selected from the group consisting of O and S; and the nucleoside analogue is a substrate for a DNA polymerase.;
14 . (canceled);
19 . A composition comprising a nucleoside analogue of the following formula: wherein R 1 is a reversible blocking group selected from the group consisting of cyanoethenyl, allenyl, cyanoethyl, formaldehyde oximyl, acrylaldehyde oximyl, propionaldehyde oximyl, and cyanoethenaldehyde oximyl; X is selected from the group consisting of O and S; and R 2 comprises a nucleobase, and optionally a linker and a detectable label or affinity agent, wherein the nucleoside analogue is covalently linked via the 5? phosphate or thiophosphate to an oligonucleotide.;
20 . (canceled);
22 . A composition comprising i) a nucleoside analogue of the following formula: or ii) a nucleoside analogue of Formula IX: wherein R 1 is a reversible blocking group selected from the group consisting of cyanoethenyl, allenyl, cyanoethyl, formaldehyde oximyl, acrylaldehyde oximyl, propionaldehyde oximyl, and cyanoethenaldehyde oximyl; X is selected from the group consisting of O and S; R 2 is a nucleobase; L is a linker; A 1 comprises an affinity tag; and A 2 comprises a detectably labeled affinity agent that forms a specific and non-covalent complex with A 1 , wherein the nucleoside analogue is covalently linked via the 5? phosphate or thiophosphate to an oligonucleotide.;
25 . (canceled);
26 . (canceled);
27 . (canceled);
28 . (canceled);
29 . (canceled);
30 . (canceled);
31 . (canceled);
32 . (canceled);
claim 21: 33 . (canceled);
claim 22: 34 . (canceled);
claim 23: 35 . (canceled);
claim 24: 36 . (canceled)

Summary

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