IP Library › Granted Patent US 12,630,808
Granted Patent B2
US 12,630,808 · App. 17/275,164 · Granted May 19, 2026

Modified archaeal family b polymerases

Inventors: Carl W. Fuller (New Jersey, NJ); Eli N. Glezer (Del Mar, CA); Andrew Spaventa (San Diego, CA); Souad Naji (La Jolla, CA)
Assignee: Singular Genomics Systems, Inc.
C12N9/1252C12P19/34C12Y207/07007
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Quick Facts
Patent No.
US 12,630,808
App. No.
17/275,164
Filed
Mar 10, 2021
Granted
May 19, 2026
Kind
B2
Art Unit
1699
USPC
435/91.2
Abstract

Provided herein are modified Archaeal family B polymerases derived from the Archaeal microorganism Pyrococcus - abyssi that exhibit improved incorporation of nucleotide analogues utilized in DNA sequencing.

Claims (30)

1 . A polymerase comprising an amino acid sequence that is at least 85% identical to a continuous 500 amino acid sequence within SEQ ID NO: 1; comprising the following amino acids:

an alanine at amino acid position 409 or at a position corresponding to amino acid position 409; and

an amino acid substitution, wherein said amino acid substitution comprises: D215A, D315A, E718A, E718N, 1522L, K602A, K712E, N735A, R705A, or R713A.

2 . The polymerase of claim 1 , wherein the polymerase comprises an amino acid sequence that is at least 95% identical to a continuous 500 amino acid sequence within SEQ ID NO: 1.

3 . The polymerase of claim 1 , wherein the polymerase comprises an alanine at amino acid position 409; an alanine or glycine at amino acid position 410; and an isoleucine, or proline at amino acid position 411.

4 . The polymerase of claim 1 , wherein the polymerase comprises an alanine at amino acid position 409; a glycine at amino acid position 410; and an isoleucine or proline at amino acid position 411.

5 . The polymerase of claim 4 , wherein the polymerase comprises an isoleucine at amino acid position 411.

6 . The polymerase of claim 1 , wherein the polymerase comprises a serine at amino acid position 409; an alanine or glycine at amino acid position 410; and a valine, isoleucine, or proline at amino acid position 411.

7 . The polymerase of claim 1 , wherein the polymerase comprises a serine at amino acid position 409; an alanine at amino acid position 410; and a valine, isoleucine, or proline at amino acid position 411.

8 . The polymerase of claim 1 , wherein the polymerase comprises an amino acid substitution mutation between position 129 and 316 of SEQ ID NO: 1, inclusive of position endpoints.

9 . The polymerase of claim 1 , wherein the polymerase comprises an amino acid substitution mutation at positions 129, 141, and 143.

10 . The polymerase of claim 1 , wherein the polymerase comprises an alanine substitution mutation at position 129.

11 . The polymerase of claim 1 , wherein the polymerase comprises an alanine substitution mutation at position 141.

12 . The polymerase of claim 1 , wherein the polymerase comprises an alanine substitution mutation at position 143.

13 . The polymerase of claim 1 , wherein the polymerase comprises an alanine substitution mutation at position 144.

14 . The polymerase of claim 1 , wherein the polymerase comprises a leucine or valine substitution mutation at position 486.

15 . The polymerase of claim 1 , wherein the polymerase comprises serine at position 515.

16 . The polymerase of claim 1 , wherein the polymerase comprises isoleucine at position 590.

17 . The polymerase of claim 1 , which exhibits an increased rate of incorporation of modified nucleotides, relative to a wild-type P. abyssi DNA polymerase.

18 . A method of incorporating a modified nucleotide into a nucleic acid sequence comprising allowing the following components to interact: (i) a DNA template, (ii) a nucleotide solution, and (iii) a polymerase, wherein the polymerase is a polymerase of claim 1 .

19 . The method of claim 18 , wherein the polymerase is capable of incorporating a modified nucleotide into a nucleic acid sequence in stringent hybridization conditions.

20 . The method of claim 18 , wherein the polymerase is capable of incorporating a modified nucleotide into a nucleic acid sequence at 55 to 80 degrees Celsius.

21 . The method of claim 18 , wherein the polymerase is capable of incorporating a modified nucleotide into a nucleic acid sequence at a pH of 8.0 to 11.0.

22 . The polymerase of claim 1 , wherein the amino acid substitution is E718A, E718N, K712E, R705A, or R713A.

23 . The polymerase of claim 1 , wherein the polymerase comprises the following amino acid substitution mutations relative to SEQ ID NO:1:

a) M129A; D141A; T144A; E143A; L409A; Y410G; P411I; A486V; T515S; K712E;

b) M129A; D141A; E143A; T144A; L409A; Y410G; P411I; A486V; T515S; T590I; R705A;

c) M129A; D141A; E143A; T144A; L409A; Y410G; P411I; A486V; T515S; T590I; R713A;

d) M129A; D141A; E143A; T144A; L409A; Y410G; P411I; A486V; T515S; T590I; E718N; or

e) M129A; D141A; E143A; T144A; L409A; Y410G; P411I; A486V; T515S; T590I; E718A.

Assignments (2)
SECURITY INTEREST Recorded Mar 7, 2025
From: SINGULAR GENOMICS SYSTEMS, INC.
To: FIRST-CITIZENS BANK & TRUST COMPANY
Reel/Frame 070440/0465 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 15, 2021
From: FULLER, CARL W.; GLEZER, ELI; SPAVENTA, ANDREW; NAJI, SOUAD
To: SINGULAR GENOMICS SYSTEMS, INC.
Reel/Frame 057490/0912 →
Continuity (2)
Provisional Application 62729875 · Sep 11, 2018
Related Publication 20230011240A1 · Jan 12, 2023
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