IP Library › Granted Patent US 12,509,669
Granted Patent B2
US 12,509,669 · App. 18/515,603 · Granted Dec 30, 2025

Engineered polymerases for improved sequencing by binding

Inventors: Pinar Iyidogan (Menlo Park, CA); Mariam Iftikhar (Menlo Park, CA); Bridget Kidd (Menlo Park, CA); Lewis Churchfield (Menlo Park, CA)
Assignee: Pacific Biosciences of California, Inc.
C12N9/1252
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Quick Facts
Patent No.
US 12,509,669
App. No.
18/515,603
Granted
Dec 30, 2025
Kind
B2
Abstract

Provided herein are engineered DNA polymerases comprising modifications improving accuracy and processivity of the polymerase including modifications in the Motif A region, optionally, along with additional modifications in the palm and/or exonuclease domains of the polymerase. Also provided are nucleic acids encoding the engineered DNA polymerases comprising modifications in motif A of the polymerase, optionally, with additional modifications. Methods, vectors, kits, and compositions comprising the nucleic acids and compositions, methods and kits comprising the engineered polymerases are also provided.

Claims (14)

1 . A method of identifying a base in a primed template nucleic acid, comprising

(a) contacting a primer-template nucleic acid hybrid with an engineered DNA polymerase and a test nucleotide complementary to a next template base, thereby forming a ternary complex, wherein the engineered DNA polymerase comprises an amino acid sequence that has at least 98% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 1, and comprises amino acid substitutions L408F, Y409F, P410T, M523T, V524T, L544F, and H545F, wherein amino acid numbering is relative to the amino acid sequence of SEQ ID NO:1;

(b) detecting the ternary complex; and

(c) identifying the next base of the primed template nucleic acid from the detected complex.

2 . The method of claim 1 , wherein, in step (b), incorporation of the test nucleotide into the primer is precluded.

3 . The method of claim 1 , wherein, in step (b), the primer comprises a blocking group at its 3′ end to preclude incorporation of the test nucleotide into the primer.

4 . The method of claim 1 , wherein the nucleotide comprises a fluorescent label that is detected in step (b) to detect the ternary complex.

5 . The method of claim 4 , the method comprising step (d) incorporating into the primer an unlabelled nucleotide that is complementary to the next template base.

6 . The method of claim 5 , wherein the unlabelled nucleotide comprises a reversible terminator.

7 . The method of claim 5 , wherein step (d) further comprises replacing the engineered DNA polymerase of steps (a) and (b) with a different type of polymerase that catalyzes the incorporation of the unlabelled nucleotide into the primer.

8 . The method of claim 5 , wherein the engineered DNA polymerase of steps (a) and (b) catalyzes the incorporation of the unlabelled nucleotide into the primer.

9 . The method of claim 1 , wherein the engineered DNA polymerase has increased processivity compared to a control polymerase without the at least one additional modification.

10 . The method of claim 9 , wherein the control polymerase comprises the amino acid sequence of SEQ ID NO:4.

11 . The method of claim 1 , wherein the engineered DNA polymerase comprises an alanine residue at position 141, an alanine residue at position 143, and a leucine residue at position 485, wherein amino acid numbering is relative to the amino acid sequence of SEQ ID NO:1.

Assignments (4)
CORRECTIVE ASSIGNMENT TO CORRECT THE CORRECT SPELLING OF THE SECOND NAMED INVENTOR'S NAME IS MARIAM IFTIKHAR PREVIOUSLY RECORDED AT REEL: 065632 FRAME: 0793. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNMENT. Recorded Dec 15, 2023
From: IYIDOGAN, PINAR; IFTIKHAR, MARIAM; KIDD, BRIDGET; CHURCHFIELD, LEWIS
To: OMNIOME, INC.
Reel/Frame 066043/0134 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 21, 2023
From: IYIDOGAN, PINAR; IFTIKHAR, MIRIAM; KIDD, BRIDGET; CHURCHFIELD, LEWIS
To: OMNIOME, INC.
Reel/Frame 065632/0793 →
MERGER Recorded Nov 21, 2023
From: OMNIOME, INC.
To: OMNIOME, LLC
Reel/Frame 065632/0936 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 21, 2023
From: OMNIOME, LLC
To: PACIFIC BIOSCIENCES OF CALIFORNIA, INC.
Reel/Frame 065633/0008 →
Continuity (4)
Division 17667379 · Feb 8, 2022
Continuation 17091998 · Nov 6, 2020
Provisional Application 62933073 · Nov 8, 2019
Related Publication 20240240162A1 · Jul 18, 2024
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