IP Library Patent Application 19208361
Patent Application
App. No. 19/208,361

HIGH FIDELITY SEQUENCING ENZYMES

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Quick Facts
Patent No.
US None
App. No.
19/208,361
Filed
May 14, 2025
Art Unit
OPAP
USPC
435/194
Abstract

Disclosed herein, inter alia, are polymerases designed for accurate incorporation of nucleotides into a primer bound to a template polynucleotide.

Claims (26)

1 . A polymerase comprising an amino acid sequence that is at least 80% identical to a continuous 500 amino acid sequence within SEQ ID NO: 1; comprising a mutation at amino acid position 306 or an amino acid position corresponding to position 306, wherein the mutation at amino acid position 306 comprises aspartic acid, glutamine, asparagine, alanine, serine, proline, valine, or glycine.

2 . The polymerase of claim 1 , wherein the mutation at amino acid position 306 is glycine, alanine, or valine.

3 . The polymerase of claim 1 , comprising leucine, isoleucine, valine, alanine, or glycine at amino acid position 341 or an amino acid position corresponding to position 341.

4 . The polymerase of claim 1 , comprising leucine, isoleucine, alanine, or glycine at amino acid position 341 or an amino acid position corresponding to position 341.

5 . The polymerase of claim 1 , comprising tyrosine, phenylalanine, tryptophan, leucine, isoleucine, or valine at amino acid position 494 or an amino acid position corresponding to position 494.

6 . The polymerase of claim 1 , comprising phenylalanine, tryptophan, leucine, isoleucine, or valine at amino acid position 494 or an amino acid position corresponding to position 494.

7 . The polymerase of claim 1 , comprising phenylalanine at amino acid position 494 or an amino acid position corresponding to position 494.

8 . The polymerase of claim 1 , comprising glutamic acid, aspartic acid, glutamine, asparagine, alanine, serine, proline, valine, or glycine at amino acid position 581 or an amino acid position corresponding to position 581.

9 . The polymerase of claim 1 , comprising tyrosine, phenylalanine, tryptophan, leucine, isoleucine, or valine at amino acid position 588 or an amino acid position corresponding to position 588.

10 . The polymerase of claim 1 , comprising lysine, arginine, histidine, glutamic acid, aspartic acid, glutamine, asparagine, alanine, serine, proline, valine, or glycine at amino acid position 280 or an amino acid position corresponding to position 280.

11 . The polymerase of claim 1 , comprising methionine, alanine, serine, leucine, isoleucine, valine, or cysteine at amino acid position 241 or an amino acid position corresponding to position 241.

12 . The polymerase of claim 1 , comprising asparagine, lysine, aspartic acid, glutamine, serine, threonine, tyrosine, or glutamic acid at amino acid position 236 or an amino acid position corresponding to position 236.

13 . The polymerase of claim 1 , comprising E306G, V341L, Y494F, E581G, and F588L.

14 . The polymerase of claim 1 , further comprising E280K, M241I, and N236D.

15 . The polymerase of claim 1 , further comprising a mutation at amino acid position 409 or an amino acid position corresponding to position 409.

16 . The polymerase of claim 15 , wherein the mutation at amino acid position 409 or the amino acid position corresponding to position 409 is alanine, glutamine, tyrosine, phenylalanine, isoleucine, valine, cysteine, serine, or histidine.

17 . The polymerase of claim 1 , comprising:

an alanine or serine at amino acid position 409 or the amino acid position corresponding to position 409; a glycine at amino acid position 410 or an amino acid position corresponding to position 410; and a proline, valine, glycine, isoleucine, or serine at amino acid position 411 or an amino acid position corresponding to position 411.

18 . The polymerase of claim 1 , further comprising at least one of the following:

a serine at amino acid position 429 or an amino acid position corresponding to position 429; a serine at amino acid position 443 or an amino acid position corresponding to position 443; a serine at amino acid position 507 or an amino acid position corresponding to position 507; and a serine at amino acid position 510 or an amino acid position corresponding to position 510.

19 . A method of incorporating a modified nucleotide into a nucleic acid sequence comprising combining in a reaction vessel: (i) a nucleic acid template, (ii) a nucleotide solution, and (iii) a polymerase, wherein the polymerase is a polymerase of claim 1 .

20 . A method of sequencing a nucleic acid sequence comprising:

a. hybridizing a nucleic acid template with a primer to form a primer-template hybridization complex;

b. contacting the primer-template hybridization complex with a DNA polymerase and modified nucleotides, wherein the DNA polymerase is the polymerase of claim 1 , wherein the modified nucleotide comprises a detectable label;

c. incorporating a modified nucleotide into the primer-template hybridization complex with the DNA polymerase to form a modified primer-template hybridization complex; and

d. detecting the detectable label; thereby sequencing a nucleic acid sequence.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 29, 2025
From: KUTCHE, BRUNA KREBS; CORPUZ, JOSHUA CATUNGAL; GLEZER, ELI N.; HANDA, SUMIT; KWAN, HENRY HOI-CHUNG; NAJI, SOUAD
To: SINGULAR GENOMICS SYSTEMS, INC.
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